Enzymatic approach of linoleic acid ruminal biohydrogenation

Ruminal biohydrogenation (BH) corresponds to a microbial reduction of dietary unsaturated fatty acid. The linoleic acid (C18:2) BH is divided into three steps: first an isomerisation into conjugated linoleic acids (CLA), then a reduction producing mainly trans-octadecenoic acids (trans-C18:1), and a final reduction producing stearic acid (C18:0). Isomerisations of CLA and trans-C18:1 can lead to a number of positional and geometrical isomers. The control of BH reactions is of interest for researchers because BH directly affects the composition of fatty acids of milk and meat. In order to better understand C18:2 BH and its variations, the development of an enzymatic approach is necessary to ascertain if the action of modulators affects the bacterial enzyme activity or ruminal bacteria. The aim of this study was to investigate the C18:2 BH capacity of ruminal content after inactivation of bacteria by chloramphenicol (Cm), an inhibitor of protein synthesis in prokaryotes. The BH of C18:2 produced mainly cis9,trans11-CLA and trans10,cis12-CLA, and trans11-C18:1 and trans10-C18:1 isomers, as previously described (Jouany et al., 2007). The increase in cis12-C18:1 came from reduction of trans10,cis12-CLA, that of trans6+7+8-C18:1 from the reduction of minor CLA isomers not quantified in this study, like trans8,trans10-CLA (Shingfield et al., 2008). The trans11 pathway was rapid: the cis9,trans11-CLA production was maximal at about 1h of incubation while trans11-C18:1 accumulated throughout incubation. On the other hand, trans10 pathway was slow: trans10,cis12-CLA regularly increased during incubation, so that it was more abundant than cis9,trans11-CLA after 3h incubation, and trans10-C18:1 only began to increase after 2h of incubation. The amount of C18:0 began to increase in the media when trans11-C18:1 concentration was over 0.05 mg/mL. Such evolution of fatty acids involved in C18:2 BH was similar to that reported in vitro with living ruminal microorganisms by Harfoot et al. (1973) and Jouany et al. (2007). So, this enzymatic approach using Cm could be an interesting and valid method to study C18:2 BH, however 3h of incubation were not sufficient to study the final reduction

Effects of fat source and dietary sodium bicarbonate plus straw on the conjugated linoleic acid content of milk of dairy cows

The effects of fat source (0.7 kg of fatty acids from extruded soybeans or palmitic acid), of sodium bicarbonate (0.3 kg) plus straw (1 kg) and the interaction of these treatments on the content of conjugated linoleic acid (CLA) in the milk of dairy cows were examined. During nine weeks a group of 10 cows received a ration with palmitic acid and bicarbonate plus straw (ration PAB). During three periods of three weeks a second group of 10 cows received successively a ration with extruded soybeans and bicarbonate plus straw (ration ESB), a ration with palmitic acid without bicarbonate or straw (ration PA), and a ration with extruded soybeans without bicarbonate or straw (ration ES). Rations ES and ESB increased the content of polyunsaturated fatty acids in milk, but decreased milk fat content, compared to rations PAB and PA. Ration ESB led to the greatest milk CLA content, by a synergy between the high amount of dietary fat, and the action of bicarbonate plus straw, favouring trans11 isomers of CLA and C18:1, presumably via a ruminal pH near neutrality. Ration ES favoured trans10 isomers, not desaturated in the mammary gland, so that the milk CLA content was lower than with ration ESB, and resulted in the lowest milk fat content. In conclusion, a ration supplemented with both extruded soybeans and bicarbonate plus straw, was an efficient way to increase the CLA content in the milk of dairy cows

Digestive anatomy and impact of dietary fibre on performances of the growing grasscutter (Thryonomys swinderianus)

The grasscutter is a rodent herbivore recently domesticated in several countries of sub-Saharan Africa for meat production. However, the development of this production to a rational and more productive model needs a better knowledge of the digestive physiology of this animal. This work aimed to study the digestive anatomy of the growing grasscutter and determine the impact of dietary fibre on growth rate and health

Effects of preconditioning and extrusion of linseed on the ruminal biohydrogenation of fatty acids. 2. In vitro and in situ studies

The extent and/or intermediates of ruminal biohydrogenation (BH) of fatty acids (FA) were investigated in vitro and in situ using a raw, pre-conditioned or extruded blend of linseed and wheat bran (70:30). The duration of in vitro incubations were 2, 4, 8, 16 and 24 h, with 5 replicates. In situ studies used 3 dry ruminally fistulated Holstein cows in a 3 × 3 Latin square design, with 3 weeks adaptation to the linseed form. The diet contained 20% (DM basis) of the linseed based blend. The duration of in situ incubations were 2, 4, 8, 16, 24 and 48 h. BH was much slower in situ than in vitro, resulting in a much lower effective disappearance of C18:2 and C18:3. Moreover, the in situ technique suggested that the technological pre-treatment of linseed did not affect C18:2 and C18:3 rate of BH, whereas reduced rates were observed in vitro. After 8 h of in vitro incubation and onwards, proportions of cis-9,trans-11C18:2 were the highest with extruded linseed. The BH of FA from linseed resulted in the appearance of great proportions of trans-10+11 to trans-16C18:1 intermediates. Extrusion increased the proportions of trans-10+11C18:1 both in vitro and in situ and proportions or trans-C18:1 were higher in situ than in vitro. Compared to previous in vivo results with the same material, the in situ method provided poor estimates of BH rates and intermediates

Polymers of triglycerides generated during heating of fat do not protect linoleic acid from ruminal biohydrogenation

Heating fats often induces a decrease of cis-9, cis-12 C18:2 and cis-9, cis-12, cis-15 C18:3 biohydrogenation (BH) in vivo (Gonthier et al. 2005), in situ (Troegeler-Meynadier et al. 2006) and in vitro (Privé et al. 2010). This is of interest because it could increase polyunsaturated fatty acids (PUFA) content of ruminant products. Temperature and duration of heating of sunflower oil affect ruminal BH of PUFA, in part due to peroxide value (Privé et al., 2010). Our hypothesis was that polymers of triglycerides (TG), formed during heating of TG but not of free FA, could be responsible for partial protection of PUFA from BH

In vitro study of dietary factors affecting the biohydrogenation shift from trans-11 to trans-10 fatty acids in the rumen of dairy cows

On the basis of the isomer-specific effects of trans fatty acids (FA) on human health, and the detrimental effect of t10,c12-conjugated linoleic acid (CLA) on cows’ milk fat production, there is a need to identify factors that affect the shift from trans-11 to trans-10 pathway during ruminal biohydrogenation of FA. This experiment was conducted in vitro and aimed at separating the effects of the diet of the donor cows from those of the fermentative substrate, which is necessary to prevent this shift. A total of four dry Holstein dairy cows were used in a 434 Latin square design. They received 12 kg of dry matter per day of four diets based on maize silage during four successive periods: the control diet (22% starch, ,3% fat); the high-starch diet, supplemented with wheat plus barley (35% starch, ,3% crude fat); the sunflower oil diet, supplemented with 5% of sunflower oil (20% starch, 7.6% crude fat); and the high-starch plus oil diet (33% starch, 7.3% crude fat). Ruminal fluid of each donor cow was incubated for 5 h with four substrates having similar chemical composition to the diets, replacing sunflower oil by pure linoleic acid (LA). The efficiency of isomerisation of LA to CLA was the highest when rumen fluids from cows receiving dietary oil were incubated with added LA. The shift from trans-11 to trans-10 isomers was induced in vitro by high-starch diets and the addition of LA. Oil supplementation to the diet of the donor cows increased this shift. Conversely, the trans-10 isomer balance was always low when no LA was added to incubation cultures. These results showed that a large accumulation of trans-10 FA was only observed with an adapted microflora, as well as an addition of non-esterified LA to the incubation substrate

The ruminal ratio of trans-10/trans-11 fatty acids obtained in vitro reflects in vivo values and strongly depends on the diet of the donor cow

Composition of fatty acids (FA) of ruminant products has a potential impact on human health. Among them, trans FA, which are intermediates of the ruminal biohydrogenation of dietary unsaturated FA, deserve interest, in particular trans-10 and trans-11 isomers, which would have negative and positive effects on human health, respectively (Tricon et al., 2004). A large variability in the ratio of trans-10 to trans-11 isomers (t10/t11) has been observed (Shingfield et al., 2006). Some dietary factors shifting from t11 to t10 have been identified, like the proportion of concentrate (Griinari et al., 1998), or the addition of oil (Roy et al., 2006). The use of in vitro systems to simulate rumen fermentation presents technical, economical and ethical advantages compared to in vivo experiments, and allows screening studies. The aim of the present study was to investigate if the ruminal fluid of a cow having the t11 to t10 deviation results in the same deviation during in vitro batch incubation and if the pathway of biohydrogenation in vitro depends mainly on the donor cow or on the fermentative substrate. This study showed that the t10/t11 ratio of the ruminal fluid after 5 hours in vitro incubation reflects the in vivo values. This ratio in vitro did not depend on cultures substrates, but related to the diet of the donor cows, suggesting a major importance of the ruminal inoculum on the biohydrogenation pathway. This might be due to the short incubation time preventing the bacterial communities to evolve according to the in vitro substrate. As a consequence, short duration batch in vitro cultures cannot be used to study the dietary conditions of the t11 to t10 shift. Nevertheless, they could possibly be used for the study of the effects of feed additives on the t10/t11 ratio

Effects of copper, zinc and selenium status on performance and health in commercial dairy and beef herds: retrospective study

A retrospective study using analysis of plasma copper and zinc, and erythrocyte glutathione peroxidase from 2 080 dairy and beef cow herds was conducted to evaluate the relationship between trace-element status and production, reproduction and health in cows and their calves. Classification of the herd status as deficient, marginal, low-adequate or high-adequate was based on the lower tercile of individual values. Odds ratios for each disorder in herds were calculated by multivariable stepwise logistic regression. Inadequate copper status was not associated with adult disorders, but was an important risk factor for poor calf performance or health. Selenium deficient status was associated with most studied disorders in cows, and both deficient and marginal herd status were strongly associated with poor health of calves, particularly with increased risks of myopathy and infectious diseases. Zinc insufficiency was strongly associated with low milk production and impaired locomotion in dairy herds, and was also associated with diarrhoea and poor growth in calves. Because a low-adequate status increased the risk of many disorders in adults and calves, we propose to classify herds as deficient and marginal when the lower terciles of plasma zinc concentration are below 12 and between 12 and 14 lmol/l respectively

Starch plus sunflower oil addition to the diet of dry dairy cows results in a trans-11 to trans-10 shift of biohydrogenation

Trans fatty acids (FA), exhibit different biological properties. Among them, cis-9,trans-11 conjugated linoleic acid has some interesting putative health properties, whereas trans-10,cis-12 conjugated linoleic acid has negative effects on cow milk fat production and would negatively affect human health. In high-yielding dairy cows, a shift from trans-11 to trans-10 pathway of biohydrogenation (BH) can occur in the rumen of cows receiving high-concentrate diets, especially when the diet is supplemented with unsaturated fat sources. To study this shift, 4 rumen-fistulated nonlactating Holstein cows were assigned to a 4 × 4 Latin square design with 4 different diets during 4 periods. Cows received 12 kg of dry matter per day of 4 diets based on corn silage during 4 successive periods: a control diet (22% starch, <3% crude fat on DM basis), a high-starch diet supplemented with wheat plus barley (35% starch, <3% crude fat), a sunflower oil diet supplemented with 5% of sunflower oil (20% starch, 7.6% crude fat), and a high-starch plus sunflower oil diet (33% starch, 7.3% crude fat). Five hours after feeding, proportions of trans-11 BH isomers greatly increased in the rumen content with the addition of sunflower oil, without change in ruminal pH compared with the control diet. Addition of starch to the control diet had no effect on BH pathways but decreased ruminal pH. The addition of a large amount of starch in association with sunflower oil increased trans-10 FA at the expense of trans-11 FA in the rumen content, revealing a trans-11 to trans-10 shift. Interestingly, with this latter diet, ruminal pH did not change compared with a single addition of starch. This trans-11 to trans-10 shift occurred progressively, after a decrease in the proportion of trans-11 FA in the rumen, suggesting that this shift could result from a dysbiosis in the rumen in favor of trans-10-producing bacteria at the expense of those producing trans-11 or a modification of bacterial activities