Effect of (+)--pinenePoster

and l-menthone on at
title text selected
70ptsButyrivibrio
species and ruminal biohydrogenation of linoleic acid in vitro
Authors 40pt text

Thulile S. Sgwane1, Sife Chikunya2 and R. John Wallace1

1Rowett Institute of Nutrition & Health, University of Aberdeen, Aberdeen, UK
2Writtle College, Lordship Road, Chelmsford, Essex, CM1 3RR

Background 60 (A) 2.0

Ruminant derived foods, meat and milk, contain a high content of saturated fat
linked to cardiovascular disease in human. Metabolism of linoleic acid (LA) by 50
ruminal bacteria of the genus Butyrivibrio transiently produces an isomer of 1.6
conjugated LA (CLA, cis-9, trans-11-18:2) which is, in contrast, thought to be
beneficial to human health. Therefore, this study investigated the possibility that 40  t-11-18:1
 LA
dietary essential oil compounds (EOC) might be useful in modifying ruminal 1.2
biohydrogenation in order to provide products with a healthier fatty acid profile.
30
 B. fibrisolvens JW11
 CLA 0.8
Methods
20
 EOC (Fig. 1) were added to M2 medium [1] to determine their effect on growth
of B. fibrisolvens JW11, B. hungatei JK615T and B. proteoclasticus P18 0.4
10
 Incubation was carried out on Hungate type-tubes at 370C and growth was
measured turbimetrically until cultures reached stationary phase. t-9,t-11-18:2
0 0.0
 Influence of (+)-α-pinene (100 mg l-1) and l-menthone (300 mg l-1) on LA 0 1 2 4 6 8 10 12 15 18 24 30 36 48
60 2.0

Fatty acid concentration (mg l-1)

metabolism by B. fibrisolvens JW11 was also investigated.
(B)
50
1.6

Optical density (nm)

40  LA
1.2
(+)--Pinene l-Menthone 30
 CLA
 JW11 0.8
Fig. 1: Structural formulae of EOC
20
t-11-18:1
Results 0.4
10
 Effects on growth varied depending on dose rate, type of EOC and bacterial sp. t-9,t-11-18:2
(Table 1). 0 0.0
 B. hungatei JK615T and B. proteoclasticus P18 sensitivity to the EOC was 0 1 2 4 6 8 10 12 15 18 24 30 36 48
60 2.0
evident from the longer lag-phase compared to B. fibrisolvens JW11 (C)
 (+)-α-Pinene showed higher antimicrobial activities than l-menthone at low dose 50
1.6
 EOC delayed disappearance of LA (Fig. 2B & C) compared to control (Fig. 2A)
and cis-9, trans-11-18:2 accumulated across treatments 40  LA
1.2
 cis-9, trans-11-18:2 reduction to trans-11-18:1 was greatly delayed by EOC thus
the B. fibrisolvens JW11 had prolonged lag-phase (Fig. 2B & C) compared to 30  JW11
the control (Fig. 2A).  CLA
t-11-18:1 0.8
20
Table 1: Effect EOC on growth of selected Butyrivibrio spp.
0.4
10
B. fibrisolvens JW11 B. hungatei JK615T B. proteoclasticus P18 t-9,t-11-18:2
Dose (mg l-1) Lag time, h Cell densitya Lag time, h Cell density Lag time, h Cell density
0 0.0
(+)--pinene
0 1 2 4 6 8 10 12 15 18 24 30 36 48
0 0 1.3±0.02 0 1.0±0.01 0 1.2±0.00
Incubation time (h)
25 7 1.3±0.06 19 1.0±0.04 17 1.2±0.01
Fig. 2: Fatty acid concentration after inoculation of B. fibrisolvens JW11
50 11 1.3±0.06 23 1.1±0.02 26 1.1±0.02 into M2 medium with 50 mg l-1 linoleic acid (LA; cis-9, cis-12-C18:2). LA
( ); cis-9, trans-11-18:2 ( ); trans-9, trans-11-18:2 ( );trans-11-18:1
75 18 1.2±0.06 31 1.0±0.01 33 1.1±0.04
( ) and bacterial growth ( ). Control [A, without EOC]; (+)--pinene
100 26 1.3±0.03 46 1.0±0.04 36 1.1±0.01 [B, 100 mg l-1]; l-menthone [C, 300 mg l-1].
l-Menthone
Conclusions
0 0 1.2±0.00 0 0.9±0.01 0 0.9±0.00
 (+)--Pinene and l-menthone had bacteriostatic effects on Butyrivibrio
200 0 1.3±0.01 0 1.0±0.01 0 0.9±0.01 spp. B. fibrisolvens JW11 was less sensitive than B. proteoclasticus
300 0 1.3±0.00 5 1.0±0.04 1 0.9±0.00 P18 and B. hungatei JK615T.

400 2 1.2±0.01 18 1.0±0.00 18 0.8±0.02  The delayed metabolism of CLA (cis-9, trans-11-18:1) in the presence
500 16 1.1±0.03 42 1.0±0.01 36 0.8±0.01 of EOC suggested inhibition of bacterial reductase enzyme systems.
Using these EOC as feed additive therefore may enhance the
600 18 1.2±0.06 52 1.0±0.01 48 0.9±0.02
availability of CLA post-ruminally.
aCell density at stationary phase. Results are mean ±SE of 3 replicates. References
1. Hobson P.N. (1969) Rumen bacteria. In: Methods in Microbiology (eds. by Norris JR & Ribbons DW),
pp. 117–32. Acad. Press., London, UK.
Acknowledgements: Thulile is a recipient of PhD studentship from Association of Commonwealth Universities

Rowett-INRA 9th Joint Symposium, Aberdeen, Scotland, UK 16 – 19 June 2014