Recent advances in chromatographic identification of CLA isomers, combined with interest in their possible properties in promoting human health (e.g., cancer prevention, decreased atherosclerosis, improved immune response) and animal performance (e.g., body composition, regulation of milk fat synthesis, milk production), has renewed interest in biohydrogenation and its regulation in the rumen. Conventional pathways of biohydrogenation traditionally ignored minor fatty acid intermediates, which led to the persistence of oversimplified pathways over the decades. Recent work is now being directed toward accounting for all possible trans-18:1 and CLA products formed, including the discovery of novel bioactive intermediates. Modern microbial genetics and molecular phylogenetic techniques for identifying and classifying microorganisms by their small-subunit rRNA gene sequences have advanced knowledge of the role and contribution of specific microbial species in the process of biohydrogenation. With new insights into the pathways of biohydrogenation now available, several attempts have been made at modeling the pathway to predict ruminal flows of unsaturated fatty acids and biohydrogenation intermediates across a range of ruminal conditions. After a brief historical account of major past accomplishments documenting biohydrogenation, this review summarizes recent advances in 4 major areas of biohydrogenation: the microorganisms involved, identification of intermediates, the biochemistry of key enzymes, and the development and testing of mathematical models to predict biohydrogenation outcomes.

Fly larvae may provide an effective method to mitigate two large and growing global concerns: the use of fish meal derived from capture fisheries in aquaculture diets and manure management in livestock and poultry facilities. A 9‐wk feed trial was conducted to determine whether fly larvae could be used as a partial fish meal and fish oil replacement in rainbow trout, Oncorhynchus mykiss, diets. A trout diet was formulated to contain 40% crude protein and 15% fat. Sixty‐seven percent of the protein in the control diet was derived from fish meal, and all the fat was derived from fish oil. Two of the test diets included using the black soldier fly, Hermetia illucens, prepupae, which are 40% protein and 30% fat, as 25 and 50% replacement for the fish meal component of the control diet. The total protein derived from black soldier fly prepupae in these two test diets was 15 and 34%, respectively. A third test diet included using housefly, Musca domestica, pupae, which is 70% protein and 16% fat, as 25% replacement for the fish meal component of the control diet. Data suggest that a rainbow trout diet where black soldier fly prepupae or housefly pupae constitute 15% of the total protein has no adverse effect on the feed conversion ratio of fish over a 9‐wk feeding period. In addition, the diet with black soldier fly prepupae permitted a 38% reduction in fish oil (i.e., from 13 to 8%); however, fish fed black soldier fly diets low in fish oil had reduced levels of omega‐3 fatty acids in their muscle fillets. The findings from this study suggest that either the black soldier fly or the housefly may be a suitable feedstuff for rainbow trout diets.

Abstract.— The black soldier fly, Hermetia illucens, has the potential to reduce animal waste on livestock facilities and produce an animal‐grade feedstuff high in protein and fat. The lipid content of insects is largely dependent on their diet. Data from this study suggest that black soldier fly prepupae incorporate α‐linolenic acid (ALA), eicosapentaenoic acid (EPA), and docosahexaenoic acid (DHA) when fish offal is included in their diet. Fly larvae were fed three different proportions of fish offal and cow manure diets over a 21‐d trial. An additional group of larvae were fed 22% fish offal diet within 24 h of their pupation. Larvae fed fish offal were, on average, 30% lipid, which was 43% more than the controls fed cow manure only, and approximately 3% of this lipid was omega‐3 fatty acids (EPA, DHA, and ALA). Furthermore, this concentration of omega‐3 fatty acids was achieved within 24 h of feeding fish offal. These omega‐3 fatty‐acid‐enhanced prepupae may be a suitable fish meal and fish oil replacement for carnivorous fish and other animal diets. In addition, they may provide a method of reducing and recycling fish offal from processing plants.

The utilization of (13)C-labeled vaccenic acid (VA) by lactating dairy cows to synthesize cis-9, trans-11 conjugated linoleic acid (CLA) was investigated. Primiparous ruminally cannulated Holstein cows (n = 3) were abomasally infused with 1.5 g of VA-1-(13)C. Blood and milk samples were taken frequently before and after VA infusion. Milk and plasma lipid were extracted using chloroform:methanol. Plasma lipid was separated into triacylglycerol (TG), cholesterol ester (CE), phospholipid (PL), nonesterified fatty acid (NEFA), and mono- and diacylglycerol (MDG) fractions. Lipid was methylated, converted to dimethyl disulfide and Diels-Alder adducts, and analyzed by GC-MS. Increased enrichment of (13)C was determined using a 2-sample t test for each sample time compared with -24 h, with significance declared at P < 0.05. Enrichment in milk fat VA was detected at 4 (3.0%), 8 (8.3%), 12 (4.1%), 16 (2.2%), and 20 h (0.8%). Enrichment in VA was also detected in plasma TG, NEFA, PL, and MDG. Enrichment in milk fat cis-9, trans-11 CLA, the Delta9-desaturase product of VA, was detected at 4 (2.6%), 8 (6.6%), 12 (3.4%), 16 (1.7%), and 24 h (0.7%). Enrichment was not detected in cis-9, trans-11 CLA for any plasma lipid fraction. Modeling of the data showed the exponential decay in (13)C enrichment over time for both VA and cis-9, trans-11 CLA in milk fat. Conversion of dietary VA to cis-9, trans-11 CLA endogenously was confirmed with the mammary gland being the primary site of Delta9-desaturase activity; approximately 80% of milk fat cis-9, trans-11 CLA originated from VA.

To determine the optimum feeding level of fatty acids of palm oil (PALM; Energizer RP10; 86.6% palmitic acid) on milk production, lactating cows (n = 18) were randomly assigned to a treatment sequence in replicated 4 x 4 Latin squares. Animals were assigned to squares by parity (3 multiparous and 1 primiparous squares with primiparous in the incomplete square). The 4 diets were designed to provide 0, 500, 1,000, and 1,500 g of PALM per day. Cows were fed individually with feed intake measured daily. Each period lasted 16 d with milk production and composition determined the final 2 d. Milk production, milk composition and feed intake data were analyzed using the MIXED procedure of SAS. Milk yields were 30.9, 34.0, 34.2, and 34.2 kg/ d (SEM = 1.9) for the 0, 500, 1,000, and 1,500 g levels, respectively. Milk yield was increased by the addition of PALM; however, there were no differences among the levels of PALM. Milk fat percentage was also increased from 3.44% for 0 g to 3.95% (SEM = 0.17) across all levels of PALM but there were no differences among the PALM treatments. Dry matter intakes were 23.3, 26.4, 24.7, and 23.8 kg/d (SEM = 1.4) for the 0, 500, 1,000 and 1,500 g levels, respectively. The addition of PALM increased milk yield and milk fat percentage, and no adverse effects on dry matter intake were observed.

A previous study showed that oleic acid was converted by mixed ruminal microbes to stearic acid and also converted to a multitude of trans octadecenoic acid isomers. This study traced the metabolism of one of these trans C18:1 isomers upon its incubation with mixed ruminal microbes. Unlabeled and labeled (18-[ 13 C] trans -9 C18:1) elaidic acid were each added to four in vitro batch cultures with three cultures inoculated with mixed ruminal bacteria and one uninoculated culture. Samples were taken at 0, 12, 24, and 48 h and analyzed for 13 C enrichment in component fatty acids by gas chromatography-mass spectrometry. At 0 h of incubation, enrichment was detected only in elaidic acid. By 48 h of incubation, 13 C enrichment was 18% ( P Ͻ 0.01) for stearic acid, 7% to 30% ( P Ͻ 0.01) for all trans C18:1 isomers having double bonds between carbons six through 16, and 5% to 10% for cis -9 and cis -11 monoenes. After 48 h, 13 C enrichment in the uninoculated cultures was only detected in the added elaidic acid. This study shows t rans fatty acids exposed to active ruminal cultures are converted to stearic acid but also undergo enzymic isomerization yielding a multitude of positional and geometric isomers. Anaerobic bacteria that colonize the rumen, or largest of the four stomach compartments in ruminant species, carry on a process of lipid biohydrogenation whereby double bonds in unsaturated fatty acids are partially or completely eliminated. Linoleic acid disappeared completely by 50 h when incubated with mixed ruminal microorganisms (1). As linoleic acid disappeared, transient increases in a number of trans diene isomers were seen, followed by the accumulation of trans -11 C18:1. During the later hours of incubation, the trans -11 C18:1 declined slowly and was accompanied by an increase in stearic acid concentration (1).Oleic acid biohydrogenation is generally presented as a direct conversion to stearic acid without the formation of trans intermediates (1, 2). When 13 C-labeled oleic acid was incubated with ruminal microorganisms in a recent study (3), enrichment was observed not only in stearic acid but also in all trans C18:1 isomers having double bonds at carbon positions six through 16. However, the fate of these positional isomers of trans -C18:1 is not clear. Trans -11 C18:1 is readily converted to stearic acid by select ruminal bacteria (4), but its conversion to other trans monenes has not been reported.Kemp et al. (5) incubated cis ( cis -2 and cis -4 to cis -13) and trans ( trans -2 and trans -5 to trans -13) octadecenoic acid isomers with a rumen Fusocillus species. They wanted to test the ability of Fusocillus to hydrogenate the octadecenoic acids to stearic acid. Cis -5 to cis -13 and trans -5 to trans -13 isomers were all hydrogenated to some extent by late log-phase cultures incubated for 3 h. Between 73% and 79% of cis -5 to cis -11 isomers were converted to stearic acid. However, cis -12 (30%) and cis -13 (5%) were poorly hydrogenated. Of the trans isomers, 45% of trans -8, trans -9, and tr...

There is limited methodology available to quantitatively assess the activity of the Delta9-desaturase enzyme in vivo without chemically inhibiting the enzyme or using radioactively labeled substrates. The objective of these experiments was to develop methodology to determine the incorporation and desaturation of 13C-labeled fatty acids into milk lipids. In a preliminary experiment, 3.7 g [1-13C]myristic acid ([1-13C]14:0), 19.5 g [1-13C]palmitic acid ([1-13C]16:0), 20.0 g [1-13C]stearic acid ([1-13C]18:0) were combined and infused into the duodenum of a cow over 24 h. In a following experiment, 5.0 g [1-13C]14:0, 40.0 g [1-13C]16:0, and 50.0 g [1-13C]18:0 were infused into the abomasums of separate cows as a bolus over 20 min or continuously over 24 h. Milk fat was extracted using chloroform:methanol. Fatty acids were methylated, and fatty acid methyl esters (FAME) were converted to dimethyl disulfide derivatives (DMDS). The FAME and DMDS were analyzed by gas chromatography mass spectrometry. In the preliminary experiment, 13C enrichment in 14:0 but not 16:0 or 18:0 was observed. When dosage amounts were increased in the following experiment, peak enrichments from the bolus infusion were observed at 8 h. Enrichments for continuous infusion peaked at 16 h for 14:0 and 18:0, and at 24 h for 16:0. The Delta9-desaturase products of these fatty acids were estimated to be 90% of cis-9 14:1, 50% of cis-9 16:1, and 59% of cis-9 18:1. This study demonstrates that 13C-labeled fatty acids may be utilized in vivo to measure the activity of the Delta9-desaturase enzyme.

Milk fat from women living in the United States contains concentrations of trans FAs similar to those in milk from Canadian women but greater than those reported in milk from women in other countries. In decreasing order of concentration, the Delta10t, Delta11t, Delta9t, and Delta12t isomers represented 78.9% of the total 18:1t. These FAs generally originate from partially hydrogenated vegetable oils and ruminant fat in the diet. No relation was found between the concentration of total trans FAs and milk fat concentration.