Background: Health-promoting polyunsaturated fatty acids (PUFA) are abundant in forages grazed by ruminants and in vegetable and fish oils used as dietary supplements, but only a small proportion of PUFA finds its way into meat and milk, because of biohydrogenation in the rumen. Butyrivibrio fibrisolvens plays a major role in this activity. The aim of this study was to investigate the mechanisms by which PUFA affect the growth of B. fibrisolvens, how PUFA are metabolized and the metabolic response to growth in the presence of PUFA. Results Linoleic acid (LA; cis-9, cis-12-18:2) and α-linolenic acid (LNA; cis-9, cis-12, cis-15-18:3) increased the lag phase of B. fibrisolvens JW11, LNA having the greater effect. Growth was initiated only when the PUFA had been converted to vaccenic acid (VA; trans- 11-18:1). The major fish oil fatty acids, eicosapentaenoic acid (EPA; 20:5(n- 3)) and docosahexaenoic acid (DHA; 22:6(n- 3)), were not metabolized and prevented growth. Cellular integrity, as determined fluorimetrically by propidium iodide (PI) ingression, was affected as much by 18:1 fatty acids, including VA, as 18:2 fatty acids. The methyl esters of LNA, LA, EPA and DHA had no effect on growth or other measurements. The ATP pool decreased by 2/3 when LA was added to growing bacteria, whereas most acyl CoA pools decreased by >96%. Conclusions It was concluded that biohydrogenation occurs to enable B. fibrisolvens to survive the bacteriostatic effects of PUFA, and that the toxicity of PUFA is probably mediated via a metabolic effect rather than disruption of membrane integrity.

This study is the first to evaluate the effects of five seaweeds (Ulva sp., Laminaria ochroleuca, Saccharina latissima, Gigartina sp., and Gracilaria vermiculophylla) on gas and methane production and ruminal fermentation parameters when incubated in vitro with two substrates (meadow hay and corn silage) for 24 h. Seaweeds led to lower gas production, with Gigartina sp. presenting the lowest value. When incubated with meadow hay, Ulva sp., Gigartina sp. and G. vermiculophylla decreased methane production, but with corn silage, methane production was only decreased by G. vermiculophylla. With meadow hay, L. ochroleuca and S. latissima promoted similar methane production as the control, but with corn silage, L. ochroleuca increased it. With the exception of S. latissima, all seaweeds promoted similar levels of total volatile fatty acid production. The highest proportion of acetic acid was produced with Ulva sp., G. vermiculophylla, and S. latissima; the highest proportion of butyric acid with the control and L. ochroleuca; and the highest proportion of iso-valeric acid with Gigartina sp. These results reveal the potential of seaweeds to mitigate ruminal methane production and the importance of the basal diet. To efficiently use seaweeds as feed ingredients with nutritional and environmental benefits, more research is required to determine the mechanisms underlying seaweed and substrate interactions.

Dietary cis-9, trans-11-conjugated linoleic acid (CLA) is generally thought to be beneficial for human health. Fish oil added to ruminant diets increases the CLA concentration of milk and meat, an increase thought to arise from alterations in ruminal biohydrogenation of unsaturated fatty acids. To investigate the mechanism for this effect, in vitro incubations were carried out with ruminal digesta and the main biohydrogenating ruminal bacterium, Butyrivibrio fibrisolvens. Linoleic acid (LA) or α-linolenic acid (LNA) was incubated (1·67g/l) with strained ruminal digesta from sheep receiving a 50:50 grass hay–concentrate ration. Adding fish oil (up to 4·17g/l) tended to decrease the initial rate of LA (P=0·025) and LNA (P=0·137) disappearance, decreased (P<0·05) the transient accumulation of conjugated isomers of both fatty acids, and increased (P<0·05) the accumulation of trans-11-18:1. Concentrations of EPA (20:5 n-3) or DHA (22:6 n-3), the major fatty acids in fish oil, were low (100mg/l or less) after incubation of fish oil with ruminal digesta. Addition of EPA or DHA (50mg/l) to pure cultures inhibited the growth and isomerase activity of B. fibrisolvens, while fish oil had no effect. In contrast, similar concentrations of EPA and DHA had no effect on biohydrogenation of LA by mixed digesta, while the addition of LA prevented metabolism of EPA and DHA. Neither EPA nor DHA was metabolised by B. fibrisolvens in pure culture. Thus, fish oil inhibits ruminal biohydrogenation by a mechanism which can be interpreted partly, but not entirely, in terms of its effects on B. fibrisolvens.