Classifications

• • A—HUMAN NECESSITIES
  • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
  • A23K—FODDER
  • A23K50/00—Feeding-stuffs specially adapted for particular animals
  • A23K50/80—Feeding-stuffs specially adapted for particular animals for aquatic animals, e.g. fish, crustaceans or molluscs
• • A—HUMAN NECESSITIES
  • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
  • A23K—FODDER
  • A23K10/00—Animal feeding-stuffs
  • A23K10/10—Animal feeding-stuffs obtained by microbiological or biochemical processes
  • A23K10/16—Addition of microorganisms or extracts thereof, e.g. single-cell proteins, to feeding-stuff compositions

Definitions

• the invention relates to feeds for aquatic organisms, such as rotifers, larval shrimp, and filter feeders.
• the feeds can be comprised of algae or yeast, or algal or yeast extracts, and can provide supplementation to the aquatic organisms, for example, with vitamins, e.g., vitamin B12, fatty acids, amino acids, growth factors, antibacterial agents, and pigments.
• DHA-enriched aquaculture feeds comprising microflora selected from Thraustochytrium, Schizochytrium, and Crypthecodinium sp., and mixtures thereof are useful for aquaculture and in particular, for feeding larval shrimp and mollusks (Barclay 2000; Gladue and Behrens 2002). Because of the small aggregate size of the algal cells, the microflora can be eaten by larval shrimp, brine shrimp, rotifers, and mollusks (Langdon et al., 1999). As an alternative approach, the present invention discloses the use of nutritionally improved yeast as aquaculture feeds.
• the improved nutritional status of yeast is obtained by selecting partially digested yeast or yeast having thin cell walls and including a mixture of certain microalgae that are used for aquaculture, such as DHA- producing microalgae and macroalgal extracts, thus formulating a complete and highly nutritious feed for aquaculture.
• These algal feeds which are based on yeast and microflora grown in fermentation medium or photoautotrophically grown algae, are suitable for use as feeds for rotifer, brine shrimp, shrimp larvae, clams, mussels, and mollusks.
• microflora grown under such conditions typically have a cell size less than about 50 ⁇ m.
• the invention provides an aquaculture feed comprising yeast and microalgae, or components thereof.
• the mean particle size can range from about 5 ⁇ m to about 100 ⁇ m, and the feed components can be dry mixed and ground to a fine powder.
• the yeast can comprise from about 30 to about 95 percent of the feed. Suitable yeast include, but are not limited to Saccharomyces spp., Saccharomyces cerevisiae, Phaffia spp., Phaffla rhodozyma, Pichia spp., Pichiapastoris, Kluyveromyces spp., Kluyveromyces aestuarii, Kluyveromyces marxianus, and Kluyveromyces yarrowii.
• the yeast can also be brewer's yeast.
• the feed can comprise at least about 5 percent by weight from microalgae. Suitable microalgae include, but are not limited to, Tetraselmis sp., Tetraselmis suecica, Myrmecia sp., Myrmecia bissecta, Lyngbya sp., Lyngbya majuscula, Cytospora sp., Scenedesmus sp., Scenedesmus obliquus, Scytonema sp., Scytonema hofinanni, Nostoc sp., Nostoc weissfloggia, Chaetoceros sp., Chaetoceros lauderi, Ecklonia sp., Ecklonia maxima, Dunaliella sp., Dunaliella salina, Dunaliella tertiolecta, Dunaliella bardiwal, Pseudoanabaena s
• Suitable microalgae also include, but are not limited to, Schizochytrium sp., Crypthecodinium sp., Crypthecodinium cohnii, Thraustochytrium sp., and components thereof.
• the feed can also comprise a probiotic element.
• Suitable probiotic elements include, but are not limited to, Lactobacillus sp., Lactobacillus casei, Lactobacillus rhamnosus, Pseudoalteromonas sp., and Pseudoalteromonas undina.
• the aquaculture feed can comprise yeast, microalgae, and macroalgae.
• the mean particle size can range from about 5 ⁇ mto about 100 ⁇ m, and the feed components can be dry mixed and ground to a fine powder.
• the yeast can comprise from about 30 to about 95 percent of the feed. Suitable yeast include, but are not limited to, Saccharomyces spp., Saccharomyces cerevisiae, Phaffla spp., Phaffia rhodozyma, Pichia spp., Pichia pastoris, Kluyveromyces spp., Kluyveromyces aestuarii, Kluyveromyces marxianus, and Kluyveromyces yarrowii. The yeast can be partially digested.
• the feed can comprise at least about 5 percent by weight from macroalgae.
• Suitable macroalgae include, but are not limited to, Tetraselmis sp., Tetraselmis suecica, Myrmecia sp., Myrmecia bissecta, Lyngbya sp., Lyngbya majuscula, Cytospora sp., Scenedesmus sp., Scenedesmus obliquus, Scytonema sp., Scytonema hofinanni, Nostoc sp., Nostoc weissfloggia, Chaetoceros sp., Chaetoceros lauderi, Ecklonia sp., Ecklonia maxima, Dunaliella sp., Dunaliella salina, Dunaliella tertiolecta, Dunaliella bardiwal, Pseudoanabaena sp., Anabaena sp.,
• the aquaculture feed can comprise at least about 0.5 percent by weight from macroalgae, which can be chosen from Laminaria spp., Padina sp., Gracillaria sp., and Ulva sp.
• the macroalgae can comprise a macroalgal extract, which can further comprise an ethanolic extraction product.
• the feed can also comprise a probiotic element. Suitable probiotic elements include, but are not limited to, Lactobacillus sp., Lactobacillus casei, Lactobacillus rhamnosus, Pseudoalteromonas sp., and Pseudoalteromonas undina. [013]
• the invention also provides a method comprising feeding a cultured species with the feed described above.
• the invention further provides a method comprising feeding a zooplankter species with a feed described above.
• Suitable zooplankter species include, but are not limited to, brine shrimp, rotifers, Artemia spp., Artemia salina,
• the purpose of the present invention is to provide an inexpensive and highly nutritious source of food derived by bacterial, algal, and/or yeast fermentation for mass production of rotifers, shrimp, larvae, and other filter feeders, such as oysters, clams, mollusks, and mussels.
• rotifers, shrimp, larvae, and other filter feeders such as oysters, clams, mollusks, and mussels.
• a major impediment to marine fish aquaculture is the high energy and labor cost (up to 85% of total operation costs according to Bolton and Fulks (Bolton 1982; Fulks and Main 1992)), and practical difficulties associated with growing high-quality algae. Frequently, it is difficult for a hatchery to supply sufficient algae at all times of the year.
• Unicellular algae are generally photoautotrophic and, as such, require light for photosynthesis during which carbon dioxide is reduced to organic carbon and oxygen is released. Hatcheries are generally using electric lighting for indoor algal cultures or, if climate conditions permit, algae can be grown in outdoor tanks. However, cell densities and growth rates are limited by light penetration into the culture medium. Photoautotrophic culture methods are energetically inefficient, costly, and require hatcheries to employ a skilled person to maintain the algal culture system. [017] It is an objective of the invention to provide rotifers, shrimp, larvae, and filter feeders with inexpensive and high quality dry feeds that are produced from phototrophically or heterotrophically grown bacteria, algae, and yeast.
• This yeast may be partially digested baker's yeast with cellulase or brewers yeast having a thin and easily digested cell wall.
• microalgae include, but are not limited to, freshwater Chlorella sp., marine Chlorella (Nannochloropsis oculata), Tetraselmis sp., Chaetoceros sp., Crypthecodinium sp., Schizochytrium sp., Chlorella spp. (such as, but not limited to, Chlorella vulgaris k-22), and Isochrysis galbana (T-iso clone).
• Crypthecodinium cohnii, and Schizochytrium sp. have exceptionally high levels of docosahexaenoic acid (DHA, 22:6n-3), which is an essential fatty acid supplement in a marine fish larval diet (reviewed by Kanazawa (Kanazawa 1993) and Watanabe (Watanabe 1993)).
• DHA docosahexaenoic acid
• Vitamin B ⁇ 2 supplemented waste stream co-products of algae up to 400 ⁇ g/100 g dry matter. More specifically, these feeds can be supplemented with additional algae-derived products containing phosphohpids rich in essential fatty acids such as docosahexaenoic (DHA, 22:6n-3) and arachidonic (ARA, 20:4n-6) acids.
• DHA docosahexaenoic
• ARA arachidonic
• These extracts can contain additional essential growth factors, antibacterial agents, bacterio static compounds, pigments, and essential amino acids.
• macroalgae include, but are not limited to, Laminaria sp., Padina sp., and Gracillaria sp.
• zooplankton refers to members of three major groups of invertebrate animals: the rotifers, copepods, and cladocerans. All three of these major groups have species adapted to pelagic (open water), littoral (vegetated), and benthic (bottom) environments, and are widespread in both freshwater and marine environments.
• Rotifer shall mean any of a class of minute, usually microscopic, but many-celled aquatic invertebrate animals having the anterior end modified into a retractile disk bearing circles of strong cilia that often give the appearance of rapidly revolving wheels. Rotifers are used as a source of live feed for aquatic animals during their early larval stage.
• Artemia belonging to the phylum Arthropoda in the class Crustacea. These tiny crustaceans can be found in salt lakes and brine ponds throughout the world. Artemia brine shrimp are used as a source of live feed for aquatic larvae during their early and late larval stages.
• aquaculture feed shall mean any dry feed in a form of fine powder that is used to culture and grow zooplanktonic animals and aquatic larvae.
• algal feed shall mean any dry feed form, which includes any portion of algal component.
• enrichment feed shall mean any type of feed either dry or moist, that is used for short duration feeding of aquatic zooplankton, prior to their harvest, in order to deliver essential nutrients for aquatic larvae when fed to these zooplankton.
• filter feeder is any species of invertebrate animal that obtains its food from aquatic media by removal of small particles or organisms by any mechanism.
• macroalgae refers to algae that in at least one life stage form large structures that are easily discernable with the naked eye. Usually these organisms have secondary vascularization and organs. Examples of different groups containing macroalgae follow but are not limited to the chlorophyta, rhodophyta, and phaeophyta.
• microalgae refers to prokaryotic and eukaryotic algae that are classed in many different species. Normally the prokaryotic algae are referred to as cyanobacteria or bluegreen algae.
• the eukaryotic microalgae come from many different genera, some of which overlap with the macroalgae and are differentiated from these by their size and a lack of defined organs (although they do have specialized cell types).
• Examples of different groups containing microalgae follow but are not limited to the chlorophyta, rhodophyta, phaeophyta, dinophyta, euglenophyta, cyanophyta, pro chlorophyta, and cryptophyta
• yeast is defined as any yeast preparation derived from yeast commonly used for production of beer, wine, or other fermented products.
• Such yeast can comprise, but are not limited to, Saccharomyces cerevisiae,
• Saccharomyces bayanus Saccharomyces bayanus, and Kluyveromyces sp.
• the rotifer and brine shrimp culture system includes fiberglass tanks at a working volume of 20-2000 L each. This system is capable of producing up to 5 x 10 9 rotifers/day/tank at a density range of 200-10,000 individuals/mL or up to 10 kg brine shrimp wet biomass/day/tank.
• the temperature is regulated in each tank by a thermostat heater at 28-32°C, depending on the rotifer strain.
• a pH controller continuously monitors the pH in each culturing tank and 10% hydrochloric acid (HCl) is added automatically by means of a Petri pump to control the pH to between 6.5-7.0.
• a buffer solution containing sodium hydroxymethane sulfonate is added to keep the ammonia in a non-toxic form. Slightly acidic pH of the culture medium is maintained, especially in high-density culture systems, since rotifer growth and fecundity rates are sensitive to high concentrations of toxic ionized ammonia (e.g., concentrations over 7.8 mg/L, as reported by Yu and Hirayama, 1986).
• Food is delivered to each culture tank manually or continuously by means of a Petri dosage pump.
• the daily amount of algal feeds and other supplements may be maintained in a chilled (0-4°C) suspension chamber, equipped with a slow mixing device.
• Rotifers are harvested continuously in a separate harvesting tank by daily exchanging of up to one third of the culture volume and can then be further enriched with essential nutrients before being delivered to fish larvae.
• a shortage of oxygen is expected. Therefore, pure oxygen, provided from oxygen cylinders or an oxygen generator, is diffused into each tank through air stones to maintain a minimal oxygen level at 3 ppm. Accumulation of parti culate organic matter and contaminating protozoans is anticipated at high culture densities, which may affect production and clog plankton nets at harvesting.
• the rotifer or brine shrimp medium is continuously circulated through nylon mat filters (1 cm thick); such a simple filtration system effectively removes solid substances with only minimal losses and stabilizes the culture performance (Yoshimura et al., 1996; Yoshimura etal, 1995).
• Brewers yeast or partially hydrolyzed baker's yeast (where at least the external layer of the cell wall is digested, either enzymatically or chemically) provide brine shrimp and rotifers with highly digestible nutrients.
• the yeast content in the algal feed formulation ranges from 30% to 95% and, more preferably from 70% to 90% of the feed.
• the present invention provides yeast that are accessible to the digestive enzymes of their predators, but their wall remains sufficiently intact so that the cell contents do not escape into the water prior to consumption.
• Dried baker's yeast is prepared using enzymatic or chemical agents, such as, but not limited to, cellulase or mannase.
• Suitable enzymes and their conditions of use are known to those skilled in the art (Lavens et al. 1992).
• Long chain polyunsaturated fatty acid-rich microalgae are provided as a source of essential fatty acids, especially DHA, ARA and EPA (Barclay and Zeller 1996; Cure et al. 1996; Furuita et al. 1998; Abu-Rezq et al. 2002; Place and Harel 2002a; Place and Harel 2002b).
• the inclusion level of DHA-rich algae ranges from 0.5 % up to 50% of the feed, and more preferably from 5% to 20% of the feed.
• DHA docosahexaenoic acid
• Larvae ultimately acquire these fatty acids from algae, either by directly feeding on algae with high levels of polyunsaturated fatty acids or by feeding on rotifers and brine shrimp that have been enriched with algae high in polyunsaturated fatty acids (Barclay and Zeller 1996).
• DHA-rich microalgae can be produced in fermentors utilizing sugar as a source of energy or photosynthetically. These algae are fortified with Vitamin B 12 , a vitamin essential for growth and reproduction of rotifers, during the production process to contain at least 200 ⁇ g/100 g dry matter.
• Arachidonic acid (ARA) rich microalgae or fungi are provided as a source of essential fatty acids.
• the inclusion level of ARA-rich algae ranges from 0.5% to 10% of the feed, and more preferably from 2% to 8% of the feed.
• ARA has been identified as an important nutrient that contributes significantly to larval growth and survival as well as helping in osmotic regulation and stress resistance.
• Probiotic or prebiotic bacteria e.g., Lactobacillus sp., Bacillus sp.,
• Bifidobacterium sp., and Enterococcus sp. are added at a range from 0.01% to 5%. These bacteria function either to colonize the gut of the organism or deliver a beneficial activity or nutrient. Such probionts could relieve stress or reduce the pathogen load of the consuming animal.
• Macroalgal extracts are also included in the formulation at a range from about 0.1 % to about 10%, and more preferably from about 0.5% to about 1%. These extracts provide the rotifers and brine shrimp with additional essential growth factors, antibacterial and bacteriostatic compounds, pigments, and essential amino acids. [042] All algal species are enriched with vitamin B 12 during production.
• the rotifer Brachionus plicatilis requires vitamin B ⁇ 2 in its diet at a level of about 100-200 ⁇ g/100 g dry matter (Hirayama et al. 1989; Hirayama and Maruyama 1991 ; Maruyama and Hirayama 1993).
• the nutritional value of the algae is further enhanced by fortifying the algae with three fat-soluble vitamins (Vitamins A, D and E) and Vitamin C. All these vitamins showed supplementary effects and significantly increased the population growth of rotifers (Satuito and Hirayama 1986; Satuito and Hirayama 1991). Examples
• Example 1 Formulation of algal feed for rotifer culture [044] Brewers dried yeast was obtained commercially from William Bio- products (Pekin, IL. USA). DHA-rich Schizochytrium sp. was obtained from Advanced BioNutrition Corp. (Columbia MD. USA). Padina sp. dry powder extract was obtained from ICP (Malta).
• Rotifer feed was formulated by mixing the ingredients in the proportions specified in Table 1. All ingredients were mixed on a dry weight basis and mill-ground to produce fine particles at sizes from about 5 to about 15 ⁇ m.
• Rotifers were fed continually with a daily ration of 0.5 g dry matter/million rotifers. Rotifers were harvested continuously by daily exchanging a total of 30-50% of the culture volume. Temperature, oxygen level, total and free ammonia, and pH were monitored continuously as outlined above. Rotifers were used directly as live feed for larval stage fish, crustaceans, or other aquatic organisms. Example 2. Formulation of algal feed for brine shrimp
• Brine shrimp start to feed about 8 h post hatching (Instar-II nauplii stage), at an initial daily ration of 100% body weight. Daily ration is reduced every day by 10 % and culture tanks can be partially harvested when the required size is t achieved. Complete harvesting can be done on day 10 post-hatch, when the brine shrimp reach maximum size or maximum biomass in the tank. Temperature, oxygen level, total and free ammonia, and pH are monitored continuously as outlined above.
• Spray-dried Spirulina sp. was obtained from Cyanotech Corp. (Kailua-
• Juvenile mussels (M. galloprovincialis) are placed in 25 L plastic aquaria at a density of 20 individuals per aquarium.
• the culture system is supplied within a closed recirculating water system and equipped with a biofilter, 10 ⁇ m mesh filter, and UV-treatment. Temperature is maintained at 24-26°C.
• Algal meal is suspended in water and vigorously mixed in a blender and fed to the mussels twice daily at a total ration of 2 mg / L. The growth rate and survival of the animals is monitored on a weekly basis.
• Example 4 Formulation of algal feed for shrimp larvae [051] Chaetoceros sp. is grown on 50% Instant Ocean medium in photobioreactors using standard lighting and procedures known in the literature (D'Souza et al. 2002). Laminaria extract is obtained from Pacific Standard Distributors hie (Menlo Park, CA). All ingredients were obtained as specified in Examples 1 and 2 and prepared according to Table 4. The dry meals were mixed and ground or milled to fine particles of less than 50 ⁇ m.
• Zooea stage shrimp larvae (Penaeus vannamei or Litopenaeus vannamei) are placed in 25 L plastic aquaria at a density of 50 individuals per aquarium.
• the culture system is supplied with a closed recirculating water system equipped with biofilter, 10 ⁇ m mesh filter and UV-treatment. The temperature is maintained between 24-26°C.
• Algal meal is suspended in water, vigorously mixed in a blender, and fed to the larvae twice daily at a total ration of 2 mg/L. The growth rate and survival of the animals are monitored on a weekly basis.
• Example 5 Formulation of algal feed for rotifer culture
• Dried brewers yeast was obtained commercially from Williams Bio- products (Pekin, IL. USA).
• DHA-rich Schizochytrium sp. was obtained from Advanced BioNutrition Corp. (Columbia MD. USA). Dry powdered Padina sp. extract was obtained from ICP Malta.
• Rotifer feed was formulated by mixing the 90 g of the brewers yeast, 9 g of the Schizochytrium biomass, and 1 g of the dried Padina extract (Table 5). All of the ingredients were mixed on a dry weight basis then mill-ground to produce fine particles at sizes between 5-15 ⁇ m.
• Rotifers were fed continually with a daily ration of 0.5 g dry matter/million rotifers. Rotifers were harvested continuously by daily exchanging a total of 30-50% of the culture volume. Temperature, oxygen level, total and free ammonia, and pH were monitored continuously. Rotifers were used directly as live feed for larval stage fish, crustaceans, and other aquatic organisms. Performance of rotifers fed on a standard culture diet (a mixture of live baker's yeast and blue green algae), a commercially available culture diet (Culture Selco), and the algal feed of the current invention is shown in Table 6.
• a standard culture diet a mixture of live baker's yeast and blue green algae
• Culture Selco commercially available culture diet
• Table 6 Production rate, doubling time, egg production and general appearance of rotifers fed different culture feeds.
• Example 6 Formulation of algal feed with probiotic Lactobacillus rhamnosus for rotifer culture
• Lactobacillus rhamnosus was obtained from Morinaga Milk Industry Co., LTD.
• Rotifer feed was formulated by mixing 89.9 g of brewers yeast, 9 g of
• Schizochytrium 1 g of Padina, and 0.1 g of L rhamnosus (Table 7). All ingredients were mixed on a dry weight basis and mill-ground to produce fine particles at sizes between about 5-15 ⁇ m.
• Rotifers were fed with a daily ration of 1 g dry matter/million rotifers.
• the culture conditions included open air tube aeration, a temperature of 28 ⁇ 1 ° C, a salinity of 20 ppt, pH 8 ⁇ 0.5, oxygen saturation 100 ⁇ 5%, and a culture volume of about 20-100 L. Daily diet rations were added 4 -5 times/day. [059] Temperature (°C), oxygen (% saturation), pH, salinity (ppt), and the number of rotifers/ml were measured daily. The daily rotifer harvest was approximately 30% of the number of rotifers in the culture. Fig.
• Example 1 shows the daily production of rotifers fed on a standard culture diet (a mixture of live baker's yeast and blue green algae), a commercially available culture diet (Culture Selco), and the algal feed with probiotic lacto bacteria of the current invention.
• a standard culture diet a mixture of live baker's yeast and blue green algae
• a commercially available culture diet Culture Selco
• Example 7 Formulation of algal feed with brewer's yeast and Schizochytrium for rotifer culture
• Dried brewers yeast was obtained commercially from Williams Bio- products (Pekin, IL. USA). DHA-rich Schizochytrium sp. was obtained from Advanced BioNutrition Corp. (Columbia MD. USA). The rotifers were grown as described in Example 6, except for the specific additions as described in Table 8. The diets containing brewers yeast and Schizochitrium at a 90:10 ratio (two different types of brewers yeast) closely matched the control diet, which was supplied with fresh algae and yeast. Additions of olive extract or Padina extract did not significantly change the number of rotifers per mL over 3 days inoculation. All of these diets bested the Culture Selco.
• Place A, R, Harel M (2002a) Use of Arachidonic acid for enhanced culturing offish larvae and broodstock.
• PCT WO 00219839, UMBI PCT WO 00219839, UMBI.
• Yoshimura K, Hgiwara A, Yoshimatsu T, Ktajima C (1996) Culture technology of marine rotifers and theimplications for intensive culture of marine fish in Japan. Mar. Freshwater Res. 47: 217-222.

Landscapes

• Life Sciences & Earth Sciences (AREA)
• Chemical & Material Sciences (AREA)
• Polymers & Plastics (AREA)
• Engineering & Computer Science (AREA)
• Animal Husbandry (AREA)
• Microbiology (AREA)
• Food Science & Technology (AREA)
• Zoology (AREA)
• Biochemistry (AREA)
• Physiology (AREA)
• Molecular Biology (AREA)
• Biotechnology (AREA)
• Biomedical Technology (AREA)
• Health & Medical Sciences (AREA)
• Insects & Arthropods (AREA)
• Marine Sciences & Fisheries (AREA)
• Birds (AREA)
• Feed For Specific Animals (AREA)
• Fodder In General (AREA)
• Farming Of Fish And Shellfish (AREA)

Applications Claiming Priority (3)

Publications (2)

Family

ID=32313100

Family Applications (1)

Country Status (4)

Families Citing this family (39)

Family Cites Families (12)

• 2003
  • 2003-11-13 AU AU2003291713A patent/AU2003291713A1/en not_active Abandoned
  • 2003-11-13 WO PCT/US2003/035043 patent/WO2004043139A2/en not_active Ceased
  • 2003-11-13 EP EP03768605A patent/EP1578191A4/de not_active Withdrawn
  • 2003-11-13 JP JP2004551696A patent/JP2006506065A/ja active Pending

Also Published As

Similar Documents

Legal Events