Food or edible material: processes – compositions – and products – Products per se – or processes of preparing or treating... – Animal derived material is an ingredient other than extract...
Reexamination Certificate
2001-03-19
2003-10-28
Sayala, Chhaya D. (Department: 1761)
Food or edible material: processes, compositions, and products
Products per se, or processes of preparing or treating...
Animal derived material is an ingredient other than extract...
C426S805000
Reexamination Certificate
active
06638561
ABSTRACT:
FIELD OF THE INVENTION
The present invention relates to the use of microbially derived arachidonic acid (ARA) for use in marine feed compositions, for feeding aquatic (e.g. marine) animals and for use in aquaculture. The ARA can be derived from a single cell organism, and thus may be in the form of a triglyceride.
BACKGROUND OF THE INVENTION
Arachidonic acid (ARA) is one of a number of Long Chain Poly-Unsaturated Fatty Acids (LC-PUFA's). Chemically it is cis-5,8,11,14 eicosatetraenoic acid (20:4) and belongs to the (n-6) family of LC-PUFA's.
Arachidonic acid is a major precursor (in fish as well as in higher terrestrial vertebrates) of a wide variety of biologically active compounds, known collectively as eicosanoids, a group comprising prostaglandins, thromboxanes and leukotrienes. Marine fish, shrimp and various terrestrial animals, such as cats, lack &Dgr;-5 desaturase activity and so are not able to convert the fatty acid 18:2(n-6) (inoleic acid) to ARA. Since arachidonic acid is essential to produce eicosanoids, it is an essential fatty acid for these animals and so must be provided in their diets.
The ratio of eicosapentaenoic acid (EPA) {20:5(n-3)} to docosahexaenoic acid DHA) {22:6(n-3)} to arachidonic acid {20:4(n-6)} in the diets of these animals can be important since a relative excess of EPA can prevent the production of eicosanoids from arachidonic acid.
Normal diets for marine fish and shrimp contain variable amounts of arachidonic acid of fish oil origin. The content of arachidonic acid in known fish oils is variable but usually relatively low, about 1% of total fatty acids present (Sargent, J. R., McEvoy, L. A. and Bell, J. G.: Aquaculture 155 (1997) p.117-127).
The ability to grow marine fish larvae is often determined to a large extent by the nutritional composition of the diet. It is difficult to obtain desirable ratios of eicosapentaenoic acid, docosahexaenoic acid and arachidonic acid using available raw materials such as fish oil, animal offal and the like since the LC-PUFA levels are highly variable even similar sources and a substantial part of arachidonic acid present is destroyed in the process of sterilising these raw materials.
One of the aims of the invention is to provide a source of arachidonic acid, of reproducible quality, to enable the aquaculture feed industry to find a satisfactory level of arachidonic acid in the diet (e.g. live feeds) for fish larvae and shrimp as well as the right ratio of arachidonic acid, eicosapentaenoic acid and docosahexanoic acid for these animals. Nearly all marine culture (mariculture) production systems rely on live feeds, principally the rotifer
Brachionus plicatilis
and nauplii of the brine shrimp Artemia, although alternative live feeds do exist.
Enrichment of live feed with eicosapentaenoic acid and docosahexaenoic acid from fish oil emulsions have been described (Sargent et al, supra). They also describe the possibility of using freeze-dried or spray-dried cells of algae and fungi containing high amounts of docosahexaenoic acid for the enrichment of live feeds.
Sargent et al also suggests arachidonic acid, but in the form of a phospholipid. Phospholipids of arachidonic acid are a natural source for flatfish and shrimp in their live feeds as this is the form that is present in fish oil. The phospholipid form of arachidonic acid is less susceptible to oxidation (in comparison with free arachidonic acid) and so is preferred by the aquaculture industry.
WO-A-90/07283 (Suntory) refers to feeds for fish and shellfish containing fungal cells which contain n-3 fatty acids (e.g. EPA, DHA) or esters thereof. However, again these sources are often low in ARA and furthermore the ARA is often present as a phospholipid.
It has been found however that by using microbially derived ARA a much higher content of ARA is achievable inside the marine animal. A marked increase in enrichment in ARA has been found in marine animals fed with feeds that contain ARA derived from single cell organisms.
DESCRIPTION OF THE INVENTION
Thus, according to a first aspect of the present invention there is provided a marine feed composition comprising one or more (marine) feed component(s) and/or ingredient(s) and microbially derived arachidonic acid (ARA). The ARA can thus be derived from a single cell organism, such as a microorganism, which it will be appreciated excludes fish oil (or marine) sources. Microbially derived ARA is usually in triglyceride form and so the invention also relates to a marine feed composition comprising ARA in the form of a triglyceride.
Preferably the feed will be (substantially) free of ARA in the form of phospholipids. The invention thus may avoid the use of phospholipid ARA, which is the form that ARA occurs in marine sources (e.g. fish oil). To thus feed marine animals with a form of ARA different from the natural source, in other words a form that would not normally be ingested by the organism, seems counter-intuitive. Nevertheless, the Applicant has found that this can lead to unexpected benefits.
The ARA will usually be present as or in an oil. Thus, this oil may be derived from or produced by a single cell organism, such as from a microbial source. The oil may have at least 10%, such as at least 20%, preferably at least 30%, advantageously at least 35% and optimally at least 40% ARA (based on the total weight of fatty acids). The oil may have an ARA content of at least 250, preferably at least 300, optimally at least 350 g/kg oil. The oil suitably has a minimum triglyceride content of 90%, eg. at least 95%, optimally at least 97%. Of the ARA present, preferably at least 85%, such as at least 90%, and optimally at least 95% is in the form of triglycerides.
Preferably the ARA is not still inside cells, for example the microbial cells that had produced the ARA. Thus, the marine feed of the invention can be (substantially) free of single cell organisms or microbes such as from which the ARA is derived or produced by. This can not only make the feed composition easier to formulate but, surprisingly, the ARA is found to be more stable once extracted from the cells that produced the ARA.
Although ARA has been suggested for use in marine feeds, the tendency has been to use ARA in a phospholipid form, usually from a marine source. This is not only because the phospholipid form is the naturally occurring form for marine animals but also since it is less susceptible to oxidation. However, despite this prejudice, the applicant has found that microbially derived ARA (or ARA in the form of a triglyceride) can provide advantages over phospholipid forms of ARA. In particular there can be a higher uptake and use of the ARA by the marine organism to which the feed composition of the invention is fed. Furthermore, with high quality microbial ARA the oxidation problem can be overcome or at least mitigated.
A further advantage of microbial ARA is that the oil can have a higher ARA content than, say, fish oil, which means less microbial oil can be used to deliver the same quantity of ARA (resulting in cheaper transport, storage and feed costs). The microbial (or single cell) source from which the ARA is derived or produced by is suitably a microorganism, for example one in a filamentous form, such as a fungus or an algae. Preferred fungi are of the order Mucorales. For example, the fungus may be of the genus Mortierella, Phycomyces, Blakeslea or Aspergillus. Preferred fungi are of the species
Mortierella alpina, Blakeslea trispora
and
Aspergillus terreus.
Preferred algae are dinoflagellates and/or belong to the genus Crypthecodinium. Preferred algae are of the species
Crypthecodinium cohnii.
These ARA sources produce predictable quantities of ARA, unlike marine sources whose PUFA content can vary according to species and the processing techniques used.
The ARA is preferably derived from or produced by the microbial (or single cell) source by fermentation. The microbes may be grown in (large scale) fermentors. Fermentation can occur when the microorganisms are present in an aqueous
Beudeker Robert Franciscus
Coutteau Peter
Morrison & Foerster / LLP
Sayala Chhaya D.
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