The objective of this study was to determine the effects of method of barley grain processing (dry-rolled vs. pelleted barley) and source of oilseed (ground canola vs. ground flaxseed), arranged as a 2 x 2 factorial, on feed intake, ruminal fermentation, nutrient flow to the duodenum, and milk production and composition in dairy cows. Eight Holstein cows (655 ± 69 kg; 83 ± 16 DIM) were used in a replicated 4 x 4 Latin square with 28-d periods. Cows in one square were fitted with ruminal and duodenal cannulae. Cows fed dry-rolled barley consumed 1.8 to 3.5 kg/d more (P = 0.02) DM than those fed pelleted barley; however, source of supplemental dietary fat had no effect on DM intake. Ruminal pH was lower (P = 0.045) in cows fed pelleted barley compared to those fed dry-rolled barley. Ruminal concentration of acetate was greater (P = 0.001), whereas ruminal concentration of propionate tended to be lower (P = 0.11), in cows fed dry-rolled barley compared to those fed pelleted barley; consequently, the acetate:propionate ratio was higher (P = 0.01) in cows fed dry-rolled barley compared to those fed pelleted barley. Ruminal concentration of total VFA was unaffected (P > 0.05) by diet. Source of dietary fat had no effect on ruminal digestion of OM, NDF, ADF or starch; however, ruminal starch digestion was slightly higher in cows fed pelleted barley compared to those fed dry-rolled barley (90.8 vs. 89.5%). Total dietary fatty acid intake was higher (P < 0.05) in cows consuming dry-rolled barley compared to those fed pelleted barley. Duodenal flow of C18:0 was lower, whereas that of C18:2n6c was higher (P < 0.05) in cows fed pelleted barley compared to those fed dry-rolled barley. Feeding flaxseed increased duodenal flows of C18:3n3, cis-9, trans-11 and trans-10, cis-12 conjugated linoleic acid compared to feeding canola. Milk yield was unaffected (P > 0.05) by diet; however, milk fat content was higher (P = 0.004) in cows fed dry-rolled barley compared to those fed pelleted barley. Milk fat content of C18:3 was higher (P = 0.005) in cows fed canola compared to those fed flax. Milk fat content of C18:3 and cis-9, trans-11 C18:2 were higher in cows fed pelleted barley compared to those fed dry-rolled barley with flax as the source of oilseed, but not with canola (interaction, P < 0.01). Milk fat content of saturated fatty acids decreased (P < 0.001) and that of polyunsaturated fatty acids increased (P = 0.003) in cows fed pelleted barley compared to those fed dry-rolled barley. In summary, milk fatty acid profiles were altered by method of grain processing and source of oilseed.

The objective of this study was to determine the effects of method of barley grain processing (dry-rolled vs. pelleted barley) and source of oilseed (ground canola vs. ground flaxseed), arranged as a 2 x 2 factorial, on feed intake, ruminal fermentation, nutrient flow to the duodenum, and milk production and composition in dairy cows. Eight Holstein cows (655 ± 69 kg; 83 ± 16 DIM) were used in a replicated 4 x 4 Latin square with 28-d periods. Cows in one square were fitted with ruminal and duodenal cannulae. Cows fed dry-rolled barley consumed 1.8 to 3.5 kg/d more (P = 0.02) DM than those fed pelleted barley; however, source of supplemental dietary fat had no effect on DM intake. Ruminal pH was lower (P = 0.045) in cows fed pelleted barley compared to those fed dry-rolled barley. Ruminal concentration of acetate was greater (P = 0.001), whereas ruminal concentration of propionate tended to be lower (P = 0.11), in cows fed dry-rolled barley compared to those fed pelleted barley; consequently, the acetate:propionate ratio was higher (P = 0.01) in cows fed dry-rolled barley compared to those fed pelleted barley. Ruminal concentration of total VFA was unaffected (P > 0.05) by diet. Source of dietary fat had no effect on ruminal digestion of OM, NDF, ADF or starch; however, ruminal starch digestion was slightly higher in cows fed pelleted barley compared to those fed dry-rolled barley (90.8 vs. 89.5%). Total dietary fatty acid intake was higher (P < 0.05) in cows consuming dry-rolled barley compared to those fed pelleted barley. Duodenal flow of C18:0 was lower, whereas that of C18:2n6c was higher (P < 0.05) in cows fed pelleted barley compared to those fed dry-rolled barley. Feeding flaxseed increased duodenal flows of C18:3n3, cis-9, trans-11 and trans-10, cis-12 conjugated linoleic acid compared to feeding canola. Milk yield was unaffected (P > 0.05) by diet; however, milk fat content was higher (P = 0.004) in cows fed dry-rolled barley compared to those fed pelleted barley. Milk fat content of C18:3 was higher (P = 0.005) in cows fed canola compared to those fed flax. Milk fat content of C18:3 and cis-9, trans-11 C18:2 were higher in cows fed pelleted barley compared to those fed dry-rolled barley with flax as the source of oilseed, but not with canola (interaction, P < 0.01). Milk fat content of saturated fatty acids decreased (P < 0.001) and that of polyunsaturated fatty acids increased (P = 0.003) in cows fed pelleted barley compared to those fed dry-rolled barley. In summary, milk fatty acid profiles were altered by method of grain processing and source of oilseed.

Oil seeds are natural sources of fat and protein in diets for lactating cows, and are usually fed whole or crushed. A recently released variety of safflower seed, 'Nutrasaff,' contains high fat (47% crude fat) and low fiber (26% NDF), and has a potential to be effectively used as a fat supplement for lactating dairy cows. Therefore, a lactating dairy cow trial was conducted to assess production performance of dairy cows when fed graded levels of whole Nutrasaff safflower seed (NSS), to determine the optimum level of NSS supplementation in the diet and to identify its impact on milk fat content and milk fatty acid (FA) profiles. Fifteen Holstein dairy cows in midlactation (118 ± 39 days in milk) were assigned into 5 groups of 3 cows each according to previous milk yield. The experimental design was a triple 5 × 5 Latin square with each period lasting 21 d (14 d of treatment adaptation and 7 d of data collection). The animals were fed a basal diet containing 56% forage (69% alfalfa hay and 31% corn silage) and 44% concentrate mix. The diet was supplemented with 0 (control), 1, 2, 3, or 4% (DM basis) whole NSS. The NSS was added to the diet by replacing whole linted-cottonseed. Intake of DM ranged from 26.4 to 27.5 kg/d across all treatments, and did not differ due to NSS inclusion. Yield of milk and ECM averaged 33.7 and 31.6 kg/d, respectively, and they were similar in response to NSS inclusion. Milk fat percentage decreased with increasing NSS inclusion, while milk protein and lactose concentrations did not differ among treatment diets. Milk fat concentration was reduced by 11% when NSS was included at 4% of the dietary DM. Feeding NSS at 1, 2, or 3% resulted in a similar milk fat concentration, and these diets also had similar milk fat percentage compared with the control diet. Concentration of milk urea N decreased by NSS inclusion regardless of level of NSS inclusion, implying that NSS supplementation improved dietary N use for milk production. Digestibilities of DM (P = 0.12) tended to increase when NSS was supplemented at 1, 2, or 3%. Cis-9, trans-11 conjugated linoleic acid (CLA) linearly increased as the NSS inclusion increased. Total concentration of n-3 FA increased by feeding NSS at 1 and 2%, whereas total concentration of n-6 FA linearly increased with increasing inclusion level of NSS. This study clearly demonstrates that it is highly possible to use NSS as a means of fat supplementation to lactating dairy cows without negative impact on lactational performance if added less than 3% of dietary DM. The enhanced milk quality with increased cis-9, trans-11 CLA concentration due to the addition of NSS could have positive implications to human health.

Increasing the milking frequency of early lactation dairy cows increases their milk yield, both during increased milking and after the cows are returned to a normal twice-daily milking schedule. When milked four times a day (4X) for only the first twenty-one days of lactation, the right udder half produces 3 kg/d more milk than the left half, which is milked only two times a day (2X) over the course of lactation (Hale et al., 2003). Alterations to this increased milking frequency (IMF) approach have been investigated in order to maximize production of the animals and determine the most efficient practice for producers. The aim of this study was to determine the appropriate duration of early lactation IMF treatment by increasing milking frequency of early lactation cows for various lengths of time, and subsequently increasing the use of this management practice on Virginia dairy farms. The right udder half of twenty-three primiparous and multiparous Holstein cows were milked 4X for 10, 20, or 40 days at the beginning of lactation, and the left udder half 2X for the entire lactation. Udder-half milk yields were measured at various time points throughout lactation and used to calculate the difference between right (4X) and left (2X) udder halves. Overall, treatment did not have a significant effect on milk yield difference throughout the entire lactation; the udder half differences for each group were -0.45 kg, 1.92 kg and 4.62 kg for the 10 d, 20 d and 40 d treatments (P > 0.05). In addition to the IMF portion of the experiment, two different methodologies were used to investigate the possible mechanism of local regulation of milk yield in response to IMF treatment. Milk fatty acid analysis was performed on milk samples obtained from the above experiment. Three different groups of fatty acids were analyzed to detect potential changes in the right udder half (4X) when compared to the left (2X); the three groups were denovo, C16, and preformed fatty acids. There was no significant effect of treatment on fatty acid composition of right and left udder halves for any of the three groups (P > 0.05). The second methodology used to explore a possible mechanism behind increased milk yield following IMF treatment was immunohistochemistry of mammary gland tissue samples obtained after IMF treatment in a previous experiment. The key target investigated was a component of the Hippo signaling pathway, Yes-associated protein (YAP). Intensity of YAP staining in the cytoplasmic area of mammary epithelial cells (MEC) and number of YAP-positive stained nuclei located in the MEC were quantified for each of the images obtained. There was no effect of treatment or day on intensity of staining (P > 0.05) with no difference in the intensity of staining between 4X and 2X samples or d 21 and d 60 samples. However, the interaction for treatment x day tended to be significant (P < 0.06), with the d 60 samples tending to have higher intensity of staining than d 21 samples. For YAP-positive nuclei, there was a significant effect of day (P < 0.05), with d 60 samples having significantly more YAP positive nuclei. There was not a significant effect of treatment or treatment x day interaction (P > 0.05) with 2X and 4X samples having the same number of YAP positive nuclei. Results from the first experiment reveal that 40 d of IMF during early lactation is sufficient to produce an increase in milk and component yields throughout lactation. This practice could be implemented on Virginia dairy farms as a way to increase efficiency and milk yield per cow. Results from the second half of this research indicates that further research is needed to investigate the fatty acid content of milk from cows subject to IMF treatment during early lactation. In addition, YAP potentially plays a role in the changes occurring in the mammary gland, with increased intensity of YAP staining and increased number of YAP positive nuclei observed at 60 DIM. Understanding of this protein and its involvement in the mammary gland could lead to identifying a mechanism for which this increase in milk yield and components following IMF is occurring. Further research needs to be done to provide results supporting the current experiment. / MSLFS

Thesis submitted in fulfilment of the requirements for the degree of Magister Technologiae: Agriculture in the Faculty of Applied Sciences at the Cape Peninsula University of Technology / Conjugated linoleic acid (CLA), a fatty acid (FA) present in the meat and milk of ruminant animals, is considered a functional nutrient for humans. The interest in CLA is attributed to its many health benefits, such as having anti-carcinogenic, anti-atherogenic, anti-diabetic and anti-adipogenic effects. Dairy products are a rich natural source of CLA, and preliminary research indicates that the levels in milk can vary based on cattle breed and feeding system. The Fleckvieh is a popular dual-purpose breed recently introduced to the Western Cape. It is used in crossbreeding programs with Holstein and Jersey herds to increase fertility and beef production. Holstein and Jersey cattle are the two most common South African dairy breeds, but little information is available on the effect of crossbreeding on FA content and CLA levels in the milk. The primary objective of this study was to compare milk FA composition and CLA content of these two dairy breeds and their respective Fleckvieh crosses. A second objective was to investigate the effect of lactation stage on CLA levels. Jersey (J) and Fleckvieh x Jersey (F×J) cows were kept in a pasture-based system (PBS) for the duration of the study. Holstein (H) and Fleckvieh x Holstein (F×H) cows were housed in a feedlot system and fed a total mixed ration (TMR) for the duration of the study. The FA and CLA content of feeds from these two feeding systems were also analysed and compared. All cows used in the study were housed at the Elsenburg Research Farm of the Western Cape Department of Agriculture. Milk samples were collected every 35 days from the cows in each feeding group, starting 10 days after calving. Samples were kept in a freezer at -20 °C until laboratory analyses. The total number of milk samples collected was 1044. The milk FA composition and CLA in particular were determined by gas chromatography. Differences between groups were statistically analysed by two-way repeated measures of analysis of variance (ANOVA). Results show that milk CLA levels are higher in Fleckvieh-crossbred cows when compared to purebred cows. The higher CLA content of milk from Fleckvieh crossbred cows demonstrates the feasibility of using Fleckvieh bulls in South African crossbreeding programs, and are also an indication that genetic selection for increased CLA content is possible. The analysis of lactation stage showed that milk CLA is higher during late lactation. A comparison of feed samples from the two feeding systems revealed that pasture feeding resulted in higher omega-3 FA in milk. Key words: Conjugated linoleic acid, milk fatty acids profile, diet, breed, health benefits, Fleckvieh, Holstein, Jersey, diet, pasture based system, total mixed ration system,

Global studies show the effect of polymorphism of selected genes on dairy production and fatty acid spectrum. The aim of the diploma thesis was genotyping of candidate FASN locus with a focus on milk yield and fatty acid spectrum depending on genotype. The thesis describes the characteristics of cow's milk, milk fat and fatty acids in milk fat. Furthermore, the thesis deals with the genome of cattle and the potential influence of polymorphism of candidate genes affecting fatty acids of milk fat. Genotypes for FASN were determined by the PCR-RFLP method, the milk yield of selected dairy cattle was statistically evaluated according to the milk production indices for 1st lactation in individual breeds, and the determination of fatty acids was performed by spectrophotometry followed by statistical evaluation. The resulting genotypes in selected breeds were only two, the GG genotype with a higher relative frequency than the genotype AG. There is no statistically significant difference between FASN genotypes depending on the milk yield and the fatty acids spectrum.

Le méthane (CH4) est un puissant gaz à effet de serre produit lors de la fermentation microbienne anaérobie des aliments dans le rumen. L’un des enjeux majeurs pour le secteur de l’élevage est de trouver des stratégies (alimentaires, génétique) pour réduire les émissions de CH4 tout en maintenant les performances animales. Les techniques de mesure de ces émissions sont coûteuses et difficilement utilisables à grande échelle sur le terrain, d’où la nécessité de trouver des alternatives de mesure ou biomarqueurs pour prédire ces émissions. Les acides gras (AG) du lait ont déjà été identifiés comme indicateurs intéressants de la méthanogenèse chez la vache laitière, mais il convient d’améliorer la précision des équations de prédiction du CH4 existantes ainsi que d'élargir leur domaine d'application à tous types de rations. L'objectif de mon travail de thèse a été de confirmer la pertinence des AG du lait comme indicateurs périphériques de la méthanogénèse chez la vache laitière avec diverses conditions nutritionnelles. Deux bases de données regroupant des données individuelles (issues d’une collaboration scientifique internationale) et moyennes (issues de la littérature) de CH4, de composition en AG du lait et d’autres performances et caractéristiques de l’animal, ainsi que des données de composition chimique des rations, ont été créées. Parallèlement, l’acquisition in vivo de données en conditions expérimentales contrôlées pour des rations mal connues ont permis d’incrémenter la base de données individuelles. Des équations de prédiction des émissions de CH4 [en g/jour, g/kg de matière sèche ingérée (MSI), et g/kg de lait] ont été développées à partir de certains AG du lait, utilisés seuls ou combinés à d’autres variables d’ingestion et de performances laitières, représentant alors des modèles complexes. Des relations entre les émissions de CH4 et la teneur de différents AG du lait (C10:0, iso C17:0 + trans-9 C16:1, iso C16:0, cis-11 C18:1, cis-15 C18:1, cis-9,cis-12 C18:2, et trans-11,cis-15 C18 :2) ont été mises en évidence, confirmant des voies métaboliques communes dans le rumen entre méthanogenèse et métabolisme lipidique. Les équations sont également liées aux types de régimes à partir desquels elles ont été développées. Les équations simples (AG du lait uniquement) sont moins précises que les complexes (erreurs résiduelles de prédiction, respectivement, de 58.6 g/jour, 2.8 g/kg MSI et 3.7 g/kg lait vs. 42.8 g/jour, 2.5 g/kg MSI et 3.3 g/kg lait). Une différence minimum de 16% de CH4 entre stratégies de réduction pourra être mise en évidence par la meilleure équation de prédiction développée. Des équations basées sur des AG bien déterminés par les méthodes infrarouges devront être testées pour évaluer, en routine et à grande échelle, de nouvelles stratégies de réduction des émissions de CH4 entérique chez la vache laitière. / Methane (CH4) is a potent greenhouse gas coming from the anaerobic microbial fermentation of the diet in the rumen. One of the main current challenge for the dairy sector is to find CH4 mitigation strategies (diets or genetics) without altering animal performance. Enteric methane measurement methods are costly and very difficult to apply on a large scale on field. Thus, there is a need to develop alternative measurement methods, such as equations based on proxies to predict CH4 emissions. Milk fatty acids (FA) have been identified as potential predictors of the methanogenesis in dairy cattle, but the prediction ability of extant published CH4 equations must be improved, and their domain of applicability must be enlarged to a wide range of diets. The objective of this PhD thesis was to confirm the potential of milk FA as proxies to predict enteric CH4 emissions in dairy cows fed a wide range of diets. Two databases (based on individual and mean data, respectively) were built thanks to an international collaboration, and gathered data on CH4, milk FA composition, dairy performances, diet and animal characteristics. Two in vivo experiments were conducted with the aim to study the effect of dietary strategies poorly documented, on methanogenesis and milk FA. The data from these experiments were included in the created database. Firstly, simple CH4 prediction equations were developed [g/d, g/kg of DMI (DMI), and g/kg of milk] based only on milk FA, and secondly other variables related to cow intake or characteristics, and dairy performance were added and constituted complex equations. Relationships between CH4 and several milk FA (C10:0, iso C17:0 + trans-9 C16:1, iso C16:0, cis-11 C18:1, cis-15 C18:1, cis-9,cis-12 C18:2, and trans-11,cis-15 C18 :2) were found, confirming common rumen metabolic pathways between methanogenesis and lipid metabolism. Equations were also closely related to the diets included in the database used for their development. Simple equations were less accurate than complex ones (prediction error of 58.6 g/d, 2.8 g/kg DMI and 3.7 g/kg milk vs 42.8 g/d, 2.5 g/kg DMI and 3.3 g/kg milk, respectively). A minimum difference of 16% in CH4 emissions between mitigating strategies can be evidenced with the best prediction equation developed in this PhD. Methane prediction equations based on milk FA well determined by infrared spectrometry methods need to be developed in order to be used on a routine basis and on a large scale. These prediction equations would allow studying the effect of novel mitigation strategies of enteric CH4 emissions in dairy cows.