Content, Pattern and Esterification of Fatty Acids in Fresh Grass in Relation to Extraction Solvents and Sample Storage Conditions

The relatively high content of linolenic acid (C18:3 n-3) in forages has lead to research on the role of forage based feeding strategies in the production of healthier milk. In these studies, forage samples are often stored, although analysis immediately after harvest has been considered to avoid oxidative deterioration and transformation of unsaturated fatty acids (FA) (Christie, 1993; Frankel, 1998). In the current study, we evaluated the effect of different storage conditions (fresh grass samples under liquid nitrogen (liq.N2) or in a cool box during 3h or at -20ºC, -80ºC or in the extraction solvent at -20°C, during 24h) on the total amount of FA extracted, the FA pattern and the extent of FA esterification. The effectiveness of isopropanol to inhibit plant enzyme activity (Hawke, 1973), which has been reported to be particularly high in plant tissues (Christie, 1993), was considered. Measures to avoid thawing losses during sample handling were also compared

Dose and time response of ruminally infused algae on rumen fermentation characteristics, biohydrogenation and Butyrivibrio group bacteria in goats

Background: Micro-algae could inhibit the complete rumen BH of dietary 18-carbon unsaturated fatty acid (UFAs). This study aimed to examine dose and time responses of algae supplementation on rumen fermentation, biohydrogenation and Butyrivibrio group bacteria in goats. Methods: Six goats were used in a repeated 3 x 3 Latin square design, and offered a fixed diet. Algae were infused through rumen cannule with 0 (Control), 6.1 (L-Alg), or 18.3 g (H-Alg) per day. Rumen contents were sampled on d 0, 3, 7, 14 and 20. Results: H-Alg reduced total volatile fatty acid concentration and acetate molar proportion (P 0.10), while H-Alg reduced the total bacteria abundance (P < 0.05). However, this was induced by a significant difference between control and H-Alg on d 14 (-4.43 %). Afterwards, both treatments did not differ as increased variation in the H-Alg repetitions, with in some cases a return of the bacterial abundance to the basal level (d 0). Conclusions: Changes in rumen fermentation and 18-carbon UFAs metabolism in response to algae were related to the supplementation level, but there was no evidence of shift in ruminal biohydrogenation pathways towards t10-18:1. L-Alg mainly induced a transient effect on rumen biohydrogenation of 18-carbon UFAs, while H-Alg showed an acute inhibition and these effects were not associated with the known hydrogenating bacteria

The economic value of information provided by milk biomarkers under different scenarios : case-study of an ex-ante analysis of fat-to-protein ratio and fatty acid profile to detect subacute ruminal acidosis in dairy cows

Monitoring systems (MS) provide additional information that many developers and researchers expect will reduce the uncertainty surrounding decision-making in livestock production and therefore enhance management decisions. However, the actual economic value of the information (VoI) yielded by MS has hardly been investigated. The aim of this study was to fill that void based on two objectives. The first is to estimate the VoI of MS prior to implementation using decision analysis based on scarce data from different sources. The second objective is to identify which factors most influence the VoI of MS and to develop recommendations about the focus of future MS development. To illustrate our objectives, we used a case study of two milk biomarkers used to monitor subacute ruminal acidosis (SARA) in dairy cows: fat-to-protein ratio (FPR) and the fatty acid profile (FAP). FPR is presently used to monitor SARA, while FAP is a newly developed test, currently in the pre-commercial phase, with reports of better accuracy than FPR. A stochastic decision tree model was used to estimate the expected monetary value of three levels of information with regards to SARA: (i) no monitoring, monitoring (ii) with FPR or (iii) with FAP. The VoI of FPR and FAP were calculated as the difference in expected monetary value of monitoring with FPR and FAP as compared with no monitoring, respectively. Several scenarios were modeled using sensitivity and elasticity analyses. The aim was not only to compensate for the scarcity of data for some variables, but also to identify under which conditions decisions based on FAP monitoring were indeed the best. In all the scenarios, monitoring SARA with FPR had the lowest expected monetary value. No monitoring was a better decision in 70% of the iterations in the scenario that described the most probable situation. The VoI of FAP was positive when SARA prevalence was between 0.21 and 0.79 with its maximum value at 0.61, when the treatment costs were lower than (sic)116/case/year and when the disease costs were higher than (sic)260/case/year. Moreover, an increase of specificity of the FAP to 0.95 yielded a positive VoI, whereas an increase of its sensitivity to 1.0 still yielded a negative VoI, suggesting that developers of the FAP should focus on improving its specificity rather than its sensitivity. To avoid suboptimal use of finite resources while developing MS, we recommend ex-ante investigation of the VoI of the MS under development

Milk fatty acid composition and associated rumen lipolysis and fatty acid hydrogenation when feeding forages from intensively managed or semi-natural grasslands

In order to evaluate the effect of replacing intensive forage by semi-natural grassland products on rumen lipid metabolism and milk fatty acid composition, four lactating and rumen canulated Holstein cows were used in a 4×4 Latin square design. Four different diets were fed: diet 100 IM - 100% intensively managed silage (IM), diet 20 SPP - 80% IM plus 20% semi-natural but species poor silage (SPP), diet 60 SPP - 40% IM plus 60% SPP and diet 60 SPR - 40% IM plus 60% semi-natural species rich silage (SPR). The silages showed significant differences in total fat content and in proportions of C18:2 n-6 and C18:3 n-3. Despite the reduced dietary supply of C18:3 n-3 with diets 60 SPP and 60 SPR, differences in milk C18:3 n-3 were small, suggesting higher recoveries of C18:3 n-3. Presumably, the latter are related to a higher transfer efficiency of C18:3 n-3 from the duodenum to the mammary gland, since rumen biohydrogenation, estimated from rumen pool size and first order rumen clearance kinetics, were similar among diets. CLA c9t11 in milk from cows fed diet 60 SPR were almost doubled compared to feeding one of the other diets. This has been related to the partial inhibition of rumen biohydrogenation of C18:3 n-3 and/or C18:2 n-6, as suggested by the increased proportions of hydrogenation isomers and reduced stearic acid proportions in rumen pool samples. In conclusion, the results suggest that the use of semi-natural grasslands in the diet of the animals reduce to some extent complete rumen biohydrogenation, which leads to an increase in milk CLA

Effect of induction of subacute ruminal acidosis on milk fat profile and rumen parameters

High-concentrate diets can lead to subacute ruminal acidosis and are known to result in changes of the ruminal fermentation pattern and mammary secretion of fatty acids. The objective of this paper is to describe modifications in milk fatty acid proportions, particularly odd- and branched-chain fatty acids and rumen biohydrogenation intermediates, associated with rumen parameters during a 6-wk subacute ruminal acidosis induction protocol with 12 ruminally fistulated multiparous cows. The protocol involved a weekly gradual replacement of a standard dairy concentrate with a wheat-based concentrate (610 g of wheat/kg of concentrate) during the first 5 wk and an increase in the total amount of concentrate in wk 6. Before the end of induction wk 6, cows were switched to a control diet because 7 cows showed signs of sickness. The pH was measured continuously by an indwelling pH probe. Milk and rumen samples were taken on d 2 and 7 of each week. Data were analyzed using a linear mixed model and by principal component analysis. A pH decrease occurred after the first concentrate switch but rumen parameters returned to the original values and remained stable until wk 5. In wk 5 and 6, rumen pH values were indicative of increasing acidotic conditions. After switching to the control diet in wk 6; rumen pH values rapidly achieved normal values. Odd- and branched-chain fatty acids and C18:1 trans-10 increased with increasing amount of concentrate in the diet, whereas C18:1 trans-11 decreased. Four fatty acids [C18:1 trans-10, C15:0 and C17:0+C17:1 cis-9 (negative loadings), and iso C14:0 (positive loading)] largely correlated with the first principal component (PC1); with cows spread along the PC1 axis. The first 4 wk of the induction experiment showed variation across the second principal component (PC2) only, with high loadings of anteiso C13:0 (negative loading) and C18:2 cis-9,trans-11 and C18:1 trans-11 (positive loadings). Weeks 5 and 6 deviated from PC2 and tended toward the negative PC1 axis. A discriminant analysis using a stepwise approach indicated the main fatty acids discriminating between the control and acidotic samples as iso C13:0, iso C16:0, and C18:2 cis-9. trans-11 rather than milk fat content. or C18:1 trans-10; which have been used before as indicators of acidosis. This shows that specific milk fatty acids have potential in discriminating acidotic cases

Identifying and exploring biohydrogenating rumen bacteria with emphasis on pathways including trans-10 intermediates

Background Bacteria involved in ruminal formation oftrans-10 intermediates are unclear. Therefore, this study aimed at identifying rumen bacteria that producetrans-10 intermediates from 18-carbon unsaturated fatty acids. Results Pure cultures of 28 rumen bacterial species were incubated individually in the presence of 40 mu g/mL 18:3n-3, 18:2n-6 ortrans-11 18:1 under control or lactate-enriched (200 mM Na lactate) conditions for 24 h. Of the 28 strains,Cutibacterium acnes(formerlyPropionibacterium acnes) was the only bacterium found to producetrans-10 intermediates from 18:3n-3 and 18:2n-6, irrespective of the growth condition. To further assess the potential importance of this species in thetrans-11 totrans-10 shift, different biomass ratios ofButyrivibrio fibrisolvens(as atrans-11 producer) andC. acneswere incubated in different growth media (control, low pH and 22:6n-3 enriched media) containing 40 mu g/mL 18:2n-6. Under control conditions, atrans-10 shift, defined in the current study astrans-10/trans-11 >= 0.9, occurred when the biomass ofC. acnesrepresented between 90 and 98% of the inoculum. A low pH or addition of 22:6n-3 inhibitedcis-9,trans-11 CLA andtrans-10,cis-12 CLA formation byB. fibrisolvensandC. acnes, respectively, wherebyC. acnesseemed to be more tolerant. This resulted in a decreased biomass ofC. acnesrequired at inoculation to induce atrans-10 shift to 50% (low pH) and 90% (22:6n-3 addition). Conclusions Among the bacterial species studied,C. acneswas the only bacterium that have the metabolic ability to producetrans-10 intermediates from 18:3n-3 and 18:2n-6. Nevertheless, this experiment revealed that it is unlikely thatC. acnesis the only or predominant species involved in thetrans-11 totrans-10 shift in vivo