MIXOTROPHIC AQUACULTURE METHOD

专利摘要:
Mixotrophic Method of Aquaculture The present invention relates to a method of culturing at least one culture organism, such as fish, shrimp or any suitable organism to be reared in an aquatic environment. An aquaculture method of at least one breeding organism is provided, the method comprising the steps: (i) providing an aquatic environment comprising at least one breeding organism, phytoplankton and bacteria; (ii) provide at least one phytoplankton nutrient and at least one bacterial nutrient during a predetermined first period, allowing phytoplankton and bacteria to grow in a first predetermined proportion of phytoplankton: bacteria greater than 1; (iii) provide at least one phytoplankton nutrient and at least one bacterial nutrient over a predetermined second period, allowing phytoplankton and bacteria to grow in a second predetermined ratio of phytoplankton: bacteria, wherein the second predetermined ratio of phytoplankton: bacteria is smaller than the first predetermined ratio of phytoplankton: bacteria; and (iv) providing at least one phytoplankton nutrient and at least one bacterial nutrient over a predetermined third period, allowing phytoplankton and bacteria to grow in a third predetermined ratio of phytoplankton: bacteria, where the third predetermined ratio of phytoplankton : bacteria is lower than the second predetermined proportion of phytoplankton: bacteria, thereby allowing at least one growing organism to grow.
公开号:BR112014031658B1
申请号:R112014031658-9
申请日:2012-06-18
公开日:2019-03-26
发明作者:Farshad Shishehchian
申请人:Blue Aqua International Pte Ltd；
IPC主号:

专利说明:

[0001] The present invention relates to an aquaculture method of at least one farmed organism, such as fish, shrimp or any organism suitable for cultivation in an aquatic environment.
Background to the Invention [0002] Aquaculture is the creation of organisms in an aquatic environment. Until the 1970s, aquaculture was not a significant contributor to the global seafood market. However, over the past 40 years, world aquaculture has expanded from an estimated 3.5 million tonnes in 1970 to around 66.7 million tonnes in 2006. In addition, government restrictions on preserving fish populations certain native species have increased the demand for seafood produced in controlled artificial environments, such as in aquaculture tanks. Catfish production on catfish farms is an example of large-scale growth in the aquaculture industry. Other species produced by the aquaculture industry include crayfish, oysters, shrimp, tilapia and striped bass.
[0003] According to the Department of Fisheries and Aquaculture of the Food and Agriculture Organization
Petition 870180161033, of 10/12/2018, p. 36/147
2/95 (FAO) of the United Nations, it is estimated that by 2012 more than 50 percent of the global consumption of food fish will come from aquaculture. With aquaculture now making a significant portion of the total seafood supply, the increase in aquaculture production has also led to significant environmental impact and competition to decrease natural resources in other sectors, such as agriculture. In particular, tank production continues to dominate aquaculture production and is especially vulnerable to water scarcity. Aquaculture farmers have therefore been under pressure to
intensify the production and grow more seafood with less water and Earth. [0004] How The production gives aquaculture if intensified over of time, provide enough oxygen
in the tank environment it has also become a major challenge. If not enough oxygen is provided, anaerobic conditions can arise and the production of toxic gas (hydrogen sulfide, ammonia) increases, affecting the health of the shrimp and thus leading to disease outbreaks. In the beginning, tank production was limited to biomass that could be sustained only with natural time-driven re-aeration. Over the years, the first emergency aeration, then the routine night aeration and finally the 24-hour aeration have been developed, which is now a
Petition 870180161033, of 10/12/2018, p. 37/147
3/95 common practice in the industry.
[0005] However, 24-hour aeration is expensive, especially in areas with limited access to electricity and / or fuel. As a general comparison, in major shrimp-producing countries such as Thailand, India or Ecuador, existing aquaculture methods reach a restocking density of 200, 100 and 30 post-larvae per square meter, respectively.
[0006] In addition, even if oxygen needs are met, concentrations of nitrogenous compounds from the decomposition of waste often reach toxic or borderline levels. The aquatic environment may also comprise organisms other than farmed organisms, such as plankton, algae and bacteria. Pathogenic or undesirable organisms can affect the growth, health and quality of cultivated species. Problems such as algae blooms and failures can also be experienced at high production rates, discouraging high stocking densities. For example, rapid growth or accumulation in populations of unwanted algae species in the aquaculture tank, in particular bluish-green algae, can result in an undesirable strange taste, causing fish meat to have an unacceptable taste and odor.
Petition 870180161033, of 10/12/2018, p. 38/147
4/95 [0007] According to FAO, China, Thailand, Vietnam, Indonesia and India dominate the world production of shrimp and river prawns. Shrimp farms can be classified as open and closed systems.
[0008] Open system shrimp farms are generally open to the environment, such as open-air tanks, built near oceans to contain and grow shrimp. These open shrimp farms suffer from the vagaries of predators, climate, disease and environmental pollution. Saltwater from the ocean must be continuously circulated through the tanks and returned to the ocean to maintain the proper water chemistry for the shrimp to grow. The
creators in shrimp should provide daily additions in tablets in dry food for custom shrimps what grow. [0009]The farms closed shrimp are
generally self-sufficient aquaculture systems. While closed shrimp farms have greater control over the artificial environment they contain, they have not been entirely satisfactory because of limited production rates, filtering problems and treatment of water and manufactured food. Although some of these deficiencies can be overcome by increased capital expenditures, such as for water treatment facilities,
Petition 870180161033, of 10/12/2018, p. 39/147
5/95 capital, labor and energy costs can be prohibitive.
[0010] Therefore, there is still a need in this technical field for improved aquaculture methods, in particular, methods that increase the intensity of production by providing high levels of oxygen and reduced levels of nitrogen compounds in the tank environment.
[0011] The material claimed here is not limited to embodiments that solve the inconveniences or that operate only in environments such as those described above. Instead, this foundation is provided only to illustrate the area of exemplary technology where some of the embodiments described here can be practiced.
Summary of the Invention [0012] The present invention solves some problems in the art and provides an aquaculture method of at least one farmed organism, wherein the farmed organism is not phytoplankton or bacteria.
[0013] According to a first aspect of the present invention, an aquaculture method of at least one breeding organism is provided, the method comprising the steps of:
[0001] provide an aquatic environment comprising at least one breeding organism, phytoplankton and bacteria;
[0002] provide at least one phytoplankton nutrient
Petition 870180161033, of 10/12/2018, p. 40/147
6/95 and at least one bacterial nutrient during a first predetermined period of time, allowing phytoplankton and bacteria to grow in a first predetermined proportion of phytoplankton: bacteria greater than 1;
[0003] providing at least one phytoplankton nutrient and at least one bacterial nutrient for a second predetermined period of time; allowing phytoplankton and bacteria to grow in a second predetermined proportion of phytoplankton: bacteria, where the second predetermined proportion of phytoplankton: bacteria is less than the first predetermined proportion of phytoplankton: bacteria; and [0004] providing at least one phytoplankton nutrient and at least one bacterial nutrient for a third predetermined period of time, allowing phytoplankton and bacteria to grow at a third predetermined proportion of phytoplankton: bacteria, where the third predetermined ratio phytoplankton: bacteria is lower than the second predetermined proportion of phytoplankton: bacteria, thus allowing at least one breeding organism to grow.
[0014] In a particular aspect, the first predetermined proportion of phytoplankton: bacteria is at
Petition 870180161033, of 10/12/2018, p. 41/147
7/95 minus about 60:40; the second predetermined proportion of phytoplankton: bacteria is between about 75:25 to about 25:75; and the third predetermined proportion of phytoplankton: bacteria is less than about 40:60.
[0015] In accordance with another aspect of the present invention, an aquaculture system capable of carrying out the method according to any aspect of the invention, the system comprising:
(a) an aquatic environment comprising breeding organisms, phytoplankton and bacteria, and / or means to provide such an environment;
(b) at least one phytoplankton nutrient providing a means to provide at least one phytoplankton nutrient to the aquatic environment;
(c) at least one phytoplankton nutrient detection means to detect at least one concentration of phytoplankton nutrients in the aquatic environment;
(d) at least one means of supplying bacterial nutrient to supply at least one bacterial nutrient to the aquatic environment;
(e) at least one bacterial addition means for adding at least one bacterium to the aquatic environment; and (f) at least one nutrient detection means for
Petition 870180161033, of 10/12/2018, p. 42/147
8/95 bacteria to detect at least one nutrient concentration of bacteria in the aquatic environment.
Brief Description of the Figures [0016] The attached figures, which are incorporated and constitute a part of this specification, illustrate aspects of the invention and, together with the general description of the invention given above and the detailed description given below, serve to explain the invention .
[0017] Figure 1 is a simplified diagram that illustrates the general nitrogen cycle in an aquatic environment, for example, in an aquaculture tank.
[0018] Figure 2 is a bar graph showing the results of the Economic Analysis done in Example 2 to compare shrimp farming using a traditional aquaculture method and the method of the present invention.
Detailed Description of the Invention [0019] It should be understood by a person skilled in the art that the present disclosure is a description of exemplary embodiments and is not intended to limit the present invention, which comprises the broader aspects embodied in the exemplary constructions.
[0020] Therefore, the present invention solves some problems in the art and provides an aquaculture method for at least one farmed organism.
Petition 870180161033, of 10/12/2018, p. 43/147
9/95 [0021] In general, the present invention is directed to an aquaculture method of at least one farmed organism. For the purposes of this specification, a breeding organism is any species commercially created or cultivated, prepared by means of aquaculture, such as any animal or plant produced by means of aquaculture, such as fish, crustaceans, molluscs, algae and / or invertebrates . Examples of types of fish include tilapia, catfish, milk fish, groupers, giant perch, carp, snakeheads, catlas, sturgeons, eels, mullets, rohus, sea bass, sparrows, rabbit fish. Examples of crustaceans include prawns, river prawns, crabs, lobsters, crayfish. Examples of mollusks include oysters, clams, mussels, scallops, clams, and sea-ears. Examples of invertebrates can include sea cucumbers, sea urchins.
[0022] For the purposes of this specification, a breeding organism can also be referred to as a primary organism or a primary breeding organism. There may be one or more breeding organisms in a given aquatic environment.
[0023] In particular, the system of the present invention is particularly well suited for raising fish and / or shrimp. Thus, most of the remaining description can be
Petition 870180161033, of 10/12/2018, p. 44/147
10/95 directed towards embodiments in which the organism created is fish and / or shrimp. It must be understood, however, that the system is also well suited for raising other aquatic farmed organisms.
[0024] In accordance with a first aspect of the present invention, an aquaculture method of at least one breeding organism is provided, the method comprising the steps:
(i) providing an aquatic environment comprising at least one breeding organism, phytoplankton and bacteria;
(ii) providing at least one phytoplankton nutrient and at least one bacterial nutrient during a first predetermined period of time, allowing phytoplankton and bacteria to grow in a first predetermined proportion of phytoplankton: bacteria greater than 1;
(iii) providing at least one phytoplankton nutrient and at least one bacterial nutrient for a second predetermined period of time; allowing phytoplankton and bacteria to grow in a second predetermined proportion of phytoplankton: bacteria, where the second predetermined proportion of phytoplankton: bacteria is less than the first predetermined proportion of phytoplankton: bacteria; and (iv) provide at least one phytoplankton nutrient
Petition 870180161033, of 10/12/2018, p. 45/147
11/95 and at least one bacterial nutrient during a third predetermined period of time, allowing phytoplankton and bacteria to grow in a third predetermined proportion of phytoplankton: bacteria, where the third
predetermined proportion in phytoplankton: bacteria is less than the second predetermined proportion in phytoplankton: bacteria, thus allowing what at least one organism in
creation grow.
[0025] Thus, it can be noted that the method of the present invention can proceed through a production cycle that can comprise at least a first, a second and a third predetermined periods. These three periods can be differentiated by the aquatic environment having:
in the first period, phytoplankton in greater abundance than bacteria;
in the second period, phytoplankton and bacteria in a smaller proportion than in the first period; and in the third period, phytoplankton and bacteria in a smaller proportion than in the second period.
[0026] In some embodiments, each of the first, second and third predetermined proportions may be greater than about 1. In other embodiments, the second and / or third predetermined proportions may be
Petition 870180161033, of 10/12/2018, p. 46/147
12/95 smaller than about 1. For example, during the third period, bacteria may be present and / or allowed to grow in greater abundance than plankton.
[0027] In a particular aspect, the first predetermined proportion of phytoplankton: bacteria is at least about 60:40; the second predetermined proportion of phytoplankton: bacteria is between about 75:25 to about 25:75; and the third predetermined proportion of phytoplankton: bacteria is less than about 40:60.
[0028]
For example, in some embodiments of the present invention, phytoplankton and bacteria can be allowed to grow in a proportion of phytoplankton:
90:10 bacteria during the first predetermined period. In such embodiments, the proportion of phytoplankton: bacteria left in the second predetermined period of time can be between about 75:25 to about
25:75. In other embodiments, the proportion of phytoplankton: bacteria left in the first predetermined period may be 60:40 and, consequently, the proportion of phytoplankton: bacteria left in the second predetermined period may be less than in the first predetermined period, i.e. , between about 60:40 and about 25:75. Likewise, the proportion of phytoplankton: bacteria left in the third period
Petition 870180161033, of 10/12/2018, p. 47/147
Predetermined 13/95 can be less than allowed in the second predetermined period of time.
[0029] In certain preferred embodiments, in the second period phytoplankton and probiotic bacteria are present and / or allowed to grow in approximately equal proportions.
[0030] The relative abundance of phytoplankton and bacteria is manipulated by regulating both inorganic matter (mineral salts and nutrients provided) and organic matter (mainly from food, but also shrimp feces and molds) in the water, including any source. of organic carbon added to the water.
[0031] However, it can be difficult or expensive to accurately determine the actual proportion of phytoplankton: bacteria. In a laboratory, phytoplankton can be counted using a microscope counting chamber, which is also able to determine which species of phytoplankton are present in the water. However, counting cell concentrations on a farm can be very time-consuming. One technique for indirectly measuring phytoplankton concentration may be to measure the amount or concentration of chlorophyll in the aquatic environment, by measuring its fluorescence. This can be done in the laboratory or using a fluorescence probe, such as probes
Petition 870180161033, of 10/12/2018, p. 48/147
14/95 aquatic plants described at https://www.ysi.com/File%20Library/Documents/Technical%20No tes / T606-The-Basics-of-Chlorophyll-Measurement.pdf, which can be implanted on or in the aquatic environment . However, the use of such probes can be very expensive, require qualified operators and / or are impractical for many aquaculture farms. Thus, many aquaculture farms typically use a Secchi disk visibility reading to estimate phytoplankton populations in the water.
[0032] As elaborated later, a Secchi disk is submerged in water and, depending on the depth (in centimeters) that disappears, the phytoplankton concentration can be estimated. An example video is available at http://www.youtube.com/watch v=yGJ5uV4jAPo. If the Secchi disk provides a measurement of 50 cm, the water has a low concentration of phytoplankton, while at 20 to 30 cm in depth, there is a high concentration of phytoplankton and nutrients should not be applied at this time, otherwise, fluorescence Excessive phytoplankton can be induced, which is harmful. The color of the water is also very important, the method of the present invention induces a brownish-green water, which may be due to the combined effect of the pigmentation of the groups of
Petition 870180161033, of 10/12/2018, p. 49/147
15/95 organisms grown.
[0033] There is also a laboratory methodology for bacterial counting, which takes even longer, requires bacterial culture and special equipment. An example of a bacterial counting protocol can be found at http://www.jochemnet.de/fiu/lab6.pdf. As the technician skilled in the subject can appreciate, bacterial populations are difficult to quantify and the data obtained depends strongly on the type and number of bacterial culture media used. In addition, this method may not be accurate, as some bacteria may not be reliably cultivable in a standard medium. Some techniques can count only the populations of one or some types of common bacteria, or types of bacteria known to be beneficial to the breeding organism (probiotic bacteria). However, bacterial culture techniques can be very time-consuming and impractical for dynamically determining the growth of bacteria and populations. There may also be more efficient high-yield techniques, such as flow cytometry with epifluorescence, but again, these techniques can be very expensive or require a lot of skill for many farms to operate profitably. Instead, on most aquaculture farms, aquaculture farmers follow environmental observations, such as the
Petition 870180161033, of 10/12/2018, p. 50/147
16/95 foam on the water surface. For example, using the method of the present invention, white foam may appear on the surface of the aquatic environment for the third predetermined period of time, possibly indicating that the bacteria are becoming more dominant. This preferably occurs at the beginning of the third predetermined period. In some embodiments, it may be possible for the foam to appear near the end of the second predetermined period of time.
[0034] In some embodiments of the invention, the steps of the invention are sequential, in which the second stage begins after the end of the first stage, the third stage after the second stage, and so on. In other embodiments of the invention, the steps of the present invention are not necessarily sequential, in which some steps can occur simultaneously with one or more other steps. For example, some embodiments may further comprise the steps of supply and / or the addition of entrants, such as minerals or bacteria. Such steps can take place over at least part of a predetermined period, preferably over more than a part of a predetermined period, more preferably, over at least two predetermined periods, even more preferably, over the first, second and third
Petition 870180161033, of 10/12/2018, p. 51/147
17/95 predetermined periods.
[0035] In some embodiments of the present invention, at least one of the phytoplankton nutrient and at least one bacterial nutrient can be provided during the first, second and third predetermined periods in the respective concentrations suitable for growing phytoplankton and bacteria in the first, second and third proportions of phytoplankton: bacteria. For example, they can be supplied at an appropriate concentration to grow phytoplankton and bacteria in a proportion of phytoplankton: bacteria from:
at least about 60:40 during the first predetermined period of time;
between about 75:25 to about 25:75 during the second predetermined period; and less than about 40:60, during the third predetermined period of time.
[0036] Different concentrations of nutrients may be necessary to promote different desired groups of phytoplankton and bacteria that are more suitable and beneficial for breeding organisms. To provide nutrients at a given concentration, an aquaculture system can be used, in which it can comprise nutrient detection means operatively coupled to the nutrient supply medium, for example, sensors to detect
Petition 870180161033, of 10/12/2018, p. 52/147
18/95 concentrations of substances contained in the water, which can indicate when additional nutrient should be supplied to the water, for example, using an automatic nutrient dispensing device.
[0037] For the purposes of this specification, the aquatic environment refers to bodies of water that serve as habitats for interrelated communities and communities and populations of plant and animal interactions, including any layer of organic matter and / or any cavity in fluid communication with the aqueous phase. For example, in a typical clay aquaculture tank, the aquatic environment includes both the aqueous phase and the solid phase that lines the bottom and sides of the tank.
[0038] Tank refers to an aquatic environment, where the species created are maintained or cultivated. In conventional fish farming, the pond is the place where young fish are bred for the size of the market. A typical tank has a clay bottom, but other materials can also be used to form the tank, for example, tanks that are concrete or plastic on the bottom are also understood to be suitable aquatic environments for the purposes of the present invention.
[0039] Aquatic environments typically also comprise organisms, such as phytoplankton and bacteria.
Petition 870180161033, of 10/12/2018, p. 53/147
19/95
When used for aquaculture, at least one breeding organism is introduced into the aquatic environment in a process known as restocking. Phytoplankton are tiny plants suspended in water with little or no ability to control their position in the water body; they can comprise microalgae and can serve as food for at least one of the breeding organism. Bacteria can comprise any form of bacteria, including bacterial spores and bacterial seeds. The bacteria may be present in aquatic environments used for aquaculture and may also be present inside or grow to colonize plants and animals in the aquatic environment.
For example, some bacteria may be present in the intestinal systems of at least one breeding organism or grow to be present in it.
Some phytoplankton or bacteria can grow to a level where they are undesirable or harmful to the health of breeding organisms, for example, by releasing certain substances that are harmful to the aquatic environment.
However, the growth of other phytoplankton and bacteria can be beneficial to the health of breeding organisms.
[0040]
For the purposes of this specification, period refers to a period of time.
[0041]
Nutrients refer to substances that are
Petition 870180161033, of 10/12/2018, p. 54/147
20/95 beneficial for the growth of an organism. For example, phytoplankton nutrients refer to substances that stimulate or promote the growth of phytoplankton, and bacterial nutrients refer to substances that stimulate or promote the growth of bacteria. Optimal growth can refer to achieving a high growth rate and / or healthy growth, so that phytoplankton and / or bacteria improve the growth of breeding organisms. For example, optimally grown phytoplankton can provide better nutrition for breeding organisms. Different organisms require different nutrients to grow, and in particular, each organism requires a different composition of nutrients for optimal growth. The nutrient composition that promotes optimal growth of green algae may be different from that required for optimal growth of blue-green algae. Likewise, different groups of bacteria grow better in environments with different nutrient compositions and different breeding organisms have different nutritional needs for optimal growth.
[0042] For the purposes of the present invention, phytoplankton and bacteria are allowed to grow for different predetermined periods in certain proportions of
Petition 870180161033, of 10/12/2018, p. 55/147
21/95 phytoplankton: bacteria. Each proportion is designed as a guide to the relative abundance of phytoplankton compared to bacteria that will provide a more beneficial environment for the growth of breeding organisms. It should be understood that the invention is not limited to growth in the exact proportions specified, since the best proportions of phytoplankton: bacteria for each predetermined period may vary depending on the breeding organism and the types of phytoplankton and bacteria present in the aquatic environment. Growing in a certain proportion can refer to growth in terms of increasing the mass of the respective organisms or in the mass of them. Growing in a certain proportion can also refer to growing in such a way that the proportion of the respective organisms is reached during the specified period. The proportions may be related to the relative abundance of phytoplankton and bacteria in terms of the mass of organisms or the number of organisms. For example, a proportion may be 60:40, which may mean that of the total number or mass of phytoplankton and bacteria, 60% may be due to phytoplankton and 40% may be due to bacteria. Growth in this proportion is achieved by, among other reasons, controlling the composition of nutrients supplied in the aquatic environment. However, phytoplankton and bacteria may not be
Petition 870180161033, of 10/12/2018, p. 56/147
22/95 present in the same proportion at all times during the predetermined period. The actual proportion of phytoplankton: bacteria present changes gradually over time, as the growth rates for each type of phytoplankton and bacteria adjust to any changes in nutrient availability and other environmental parameters, for example, dissolved oxygen, temperature and intensity of sunlight. Both the growth of organisms and the numbers in which they are present can be tedious to establish directly, therefore, for the purposes of this specification, they can be measured indirectly, for example, by means of water visibility readings, concentrations of their metabolites or consumption of resources, for example, change in dissolved oxygen, and the like.
[0043] In some embodiments of the present invention, at least one phytoplankton nutrient can be provided during the first, second and third predetermined periods in decreasing concentrations suitable for growing phytoplankton and bacteria in the first, second and third proportions of phytoplankton: predetermined bacteria. For example, phytoplankton nutrients can be supplied in decreasing concentrations suitable for growing phytoplankton and bacteria in a proportion of
Petition 870180161033, of 10/12/2018, p. 57/147
23/95 phytoplankton: bacteria from:
at least about 60:40 during the first predetermined period of time;
between about 75:25 to about 25:75 during the second predetermined period; and less than about 40:60 during the third predetermined period of time. The concentration of phytoplankton nutrients can be gradually reduced over the first, second and third predetermined periods in any combination of one step, linear and / or exponential decrease. In addition, the reduction can be moderated dynamically in response to certain measurable parameters, such as, but not limited to, dissolved oxygen, dissolved nitrogen compounds, compounds containing dissolved phosphorus and the visibility of the Secchi disk. For example, if the visibility of the Secchi disk indicates that phytoplankton is not growing sufficiently to achieve the desired phytoplankton: bacteria ratio, more phytoplankton nutrients can be provided, increasing the concentration of phytoplankton nutrients and promoting phytoplankton growth, so that phytoplankton and bacteria are allowed to grow in the desired proportion. In addition, the reduction can be mitigated in response to certain easily observable indicators, however
Petition 870180161033, of 10/12/2018, p. 58/147
24/95 less easily measurable, such as the color of the water in the aquatic environment. Under normal growing conditions when using the method of the present invention, the water in the aquatic environment can preferably be green, brown, light brown or brownish green. Preferably, the water is brownish green in color. However, in some cases, green water or other water colors may indicate a bloom of harmful phytoplankton algae, such as blue-green algae, and the decrease in phytoplankton nutrient concentration can be moderated to alleviate this. If desired, it may also be possible in some embodiments to use at least one means of detecting bacteria, such as an apparatus capable of counting and identifying the various types of bacteria. Such an apparatus can, for example, comprise a genetic analysis device such as that provided for at http://www.springerlink.com/content/v5443m2823833888/. However, in most cases, the use of such devices may not currently be possible on a large scale due to cost reasons.
[0044] In some embodiments of the present invention, at least one bacterial nutrient can be provided during the first, second and third predetermined periods in increasing concentrations suitable for
Petition 870180161033, of 10/12/2018, p. 59/147
25/95 grow phytoplankton and bacteria in the first, second and third proportions of phytoplankton: predetermined bacteria. For example, nutrients from bacteria can be supplied in increasing concentrations suitable for growing phytoplankton and bacteria in a proportion of phytoplankton: bacteria from:
at least about 60:40 during the first predetermined period of time;
between about 75:25 to about 25:75 during the second predetermined period; and less than about 40:60 during the third predetermined period of time. The nutrient concentration of bacteria can be gradually increased over the first, second and third predetermined periods, in any combination of one-step, linear and / or exponential increase. The increase can also be moderated dynamically in response to certain measurable parameters, such as, but not limited to, dissolved oxygen, dissolved nitrogen compounds and dissolved organic carbon. In addition, the increase can be modulated in response to certain easily observable but less easily measurable indicators, such as the color and appearance of foam on the surface of the aquatic environment. Under normal growth conditions when using the method
Petition 870180161033, of 10/12/2018, p. 60/147
26/95 of the present invention, foam may appear during the third predetermined period of time, which may indicate that populations of bacteria are becoming prevalent in the aquatic environment. Preferably, the foam may be white in color. Therefore, to give an example, if the expected occurrence of foam does not develop in the third predetermined period of time, this may indicate that the bacteria are not growing fast enough to become dominant, and therefore an aquaculturist may decide to provide more nutrients bacteria, increasing the nutrient concentration of bacteria and promoting the growth of bacteria, so that phytoplankton and bacteria are allowed to grow in the desired proportion.
[0045] In some embodiments of the present invention, the method may further comprise adding bacteria in the aquatic environment, wherein the added bacteria are able to maintain the concentration of ammonia and / or nitrites and / or nitrates in the aquatic environment at a level that is non-toxic to at least one breeding organism, and / or where the bacteria is non-toxic or apatogenic to at least one breeding organism. Bacteria can be added during the first, second and third predetermined periods in appropriate increased concentrations to allow phytoplankton and bacteria to grow in
Petition 870180161033, of 10/12/2018, p. 61/147
27/95 first, second and third predetermined proportions of phytoplankton: bacteria. For example, bacteria can be added to increase appropriate concentrations to allow phytoplankton and bacteria to grow in a proportion of phytoplankton: bacteria from:
at least about 60:40 during the first predetermined period of time;
between about 75:25 to about 25:75 during the second predetermined period; and less than about 40:60 during the third predetermined period of time. As with embodiments where the nutrient concentration of bacteria is gradually increased, the increase in added bacteria can also be moderated dynamically in response to certain measurable parameters, and / or certain easily observable but less easily measurable indicators, such as color and the appearance of foam on the surface of the aquatic environment.
[0046] The invention manages the sediment and the soil of the tank, and improves the quality of water and effluents, minimizing the environmental impact. The breeding organism's survival rate is improved and the risk of failure due to low production or disease is minimized, and less chemical compounds are needed. These benefits come in part from
Petition 870180161033, of 10/12/2018, p. 62/147
28/95 due to beneficial bacteria that reduce the accumulation of decomposing organic matter, thus preventing the excessive increase in biological oxygen demand (BOD), while simultaneously competing and suppressing growth in populations of toxic bacteria and / or harmful.
Consequently, it can be understood by a technician skilled in aquaculture that a gradually decreasing concentration of phytoplankton nutrients is provided to have more phytoplankton during the first predetermined period and to gradually decrease its concentration in the second and third predetermined periods. A gradually increasing concentration of nutrient bacteria is provided and a gradually increasing amount of bacteria is added to increase its population gradually over the first, second and third predetermined periods. Thus, over this time, there is a shift from a predominance of phytoplankton to bacteria.
[0048] The desired proportions for each period depending on the organism's phytoplankton: predetermined bacteria can vary from breeding and the types of phytoplankton and bacteria present in the aquatic environment. In some embodiments of the present invention, phytoplankton and bacteria can be allowed to grow in a first
Petition 870180161033, of 10/12/2018, p. 63/147
29/95 predetermined proportion of phytoplankton: bacteria of at least about 65:35 during the first predetermined period. In particular embodiments, this ratio may be at least about 70:30, preferably at least about 75:25, more preferably at least about 80:20. In even more preferred embodiments, this ratio can be at least about 85:15, more preferably 90:10. In some particular embodiments, this ratio can be about 95: 5.
[0049] In some embodiments of the present invention, phytoplankton and bacteria can be grown in a second proportion of phytoplankton: bacteria from about 70:30 to about 30:70 during the second predetermined period. More preferably, this ratio can be between about 65:35 and about 35:65, even more preferably about 60:40 to about 40:60. In particular preferred embodiments, the ratio can be between about 55:45 and about 45:55. In some particular preferred embodiments, phytoplankton and bacteria can be equally dominant, that is, this ratio can be about 50:50.
[0050] In some embodiments of the present invention, the third predetermined proportion may be less
Petition 870180161033, of 10/12/2018, p. 64/147
30/95 than the second predetermined ratio, which can be less than the first predetermined ratio, which can be greater than 1. For example, the first, second and third predetermined ratios can be 90:10, 75:25 and 50:50, respectively. In another example, the first, second and third predetermined proportions can be 75:25, 50:50 and 25:75, respectively.
[0051] Aquaculture methods known in the art basically control the growth of phytoplankton, even if probiotics are applied, growers do not follow any protocol to maintain bacterial populations. Only phytoplankton are promoted to grow in the traditional shrimp farming technique.
[0052] aquaculture methods known in the art also maintain the growth of phytoplankton in levels resulting in readings from the Secchi disk from 30 to 35 cm. This can be as more phytoplankton is perceived to represent more abundant food for breeding organisms, and is therefore considered beneficial. However, the inventors have found that this can lead to problems that may be due to the increased biological oxygen demand from the decomposition of dead plankton and hypoxia conditions for the breeding organism.
[0053] Often the excessive flowering of
Petition 870180161033, of 10/12/2018, p. 65/147
31/95 phytoplankton leads to the fall of phytoplankton and consequent anaerobic conditions. There is a high demand for oxygen from bacteria when phytoplankton fail, as they break down dead phytoplankton. If sufficient oxygen is not provided, anaerobic conditions can arise and the production of toxic gas (hydrogen sulfide, ammonia) increases, affecting the health of the shrimp and thus leading to disease outbreaks.
[0054] Instead, the inventors have surprisingly found that having less phytoplankton and reducing populations of phytoplankton in relation to the population of bacteria can be beneficial for breeding organisms. Therefore, in some embodiments of the present invention, the phytoplankton can be allowed to grow so that the aquatic environment has a visibility of the Secchi disk between about 60 cm and about 30 cm during the first predetermined period;
the aquatic environment has a visibility of the Secchi disk between about 40 cm and about 20 cm during the second predetermined period; and the aquatic environment has a visibility of the Secchi disk of between about 70 cm and about 60 cm during the third predetermined period.
[0055] Secchi Disk Visibility: The test of
Petition 870180161033, of 10/12/2018, p. 66/147
32/95 visibility of the Secchi disk is a commonly used measure of water quality and plankton abundance in the aquaculture tank. A standard Secchi disk is a 20 cm diameter disk with black and white quadrants
alternate. It is linked to an line in calibration and equipped with a Weight, so what sink quickly. At the point at which the disco from Secchi fades away of sight, O
line length from the water surface to the top of the Secchi disk is measured. This is the visibility of Secchi's Disc. The visibility of the Secchi disk is usually indicated in centimeters and can vary widely from a few centimeters to several meters. Generally, there is not enough light for the plants to grow to about twice the visibility of the Secchi disk. Thus, twice the visibility of the Secchi disk is a rough estimate of the depth of the photic zone in lakes, lagoons and other bodies of water.
[0056] The visibility of the Secchi disk is strongly related to water turbidity and can be affected by particles suspended from soil sediments. In this regard, the technician versed in the subject must take into account the visibility of the previous Secchi disk to assess the turbidity of the water, due to the abundance of plankton. The test is commonly used in aquaculture to assess abundance of
Petition 870180161033, of 10/12/2018, p. 67/147
33/95 plankton and as an indicator of the need to apply fertilizers in fish or shrimp culture to encourage plankton growth. Changes in the visibility of Secchi's disc over time are also important to indicate changes in plankton abundance.
The passage of light through a column of water is described by the equation:
Light in depth z = Incident light x and ^_kz where e = base of the natural logarithm (2.303), k = extinction coefficient, ez = depth in meters.
[0057] It has been shown that the extinction coefficient is closely related to the visibility of Secchi's disk in meters: k = 1.7 / visibility of Secchi's disk in meters.
[0058] Since the visibility of the Secchi disk is used to compare clarity between aquatic organisms, a standard procedure must be followed for its measurement, or serious errors of interpretation may occur. Guidelines for properly measuring the visibility of the Secchi disk include, for example:
• The disc must be lowered slowly until it disappears only after viewing a first measurement taken. Then it should be lifted until it only
Petition 870180161033, of 10/12/2018, p. 68/147
34/95 reappear. The average of the two measurements should be used according to the visibility of the Secchi disk.
• The measurement should be made on clear or partially cloudy days, when the sun is not obscured by clouds, and the reading must be made with the sun behind the observer.
• The observer's face must be within 25 to 50 cm above the water surface when taking the reading, and the observer should not wear sunglasses while taking the measurement.
[0059] In some embodiments of the present invention, the method may further comprise the step of providing at least one additional feed for at least one breeding organism to grow, the additional feed being provided in a 1: A: B ratio in the first , second and third predetermined periods, respectively, where A is between about 3 and about 15 and / or B is between about 10 and about 30. In particular embodiments, A can be between about 5 and about 10 and / or B can be between about 15 and about
20. The aquaculturist versed in the subject would understand that the proportions of additional food to be provided can refer to an accumulated amount or a dose rate, and that both the accumulated amount and the dose rate of the
Petition 870180161033, of 10/12/2018, p. 69/147
35/95 additional feed will depend on the species of breeding organisms and the stocking density. For example, in shrimp farming a restocking density of at least about 200 post-larvae per square meter of tank size, at least one of the additional feed can be provided at a daily rate of between about 5 kg to about 15 kg during the first predetermined period, at a daily rate of between about 15 kg to about 75 kg during the second predetermined period, and between about 50 kg to about 150 kg during the third predetermined period. If the first, second and third predetermined periods are of equal size, the cumulative amounts of additional feed would be in ratios similar to the ratio between dosage rates. However, if the predetermined periods are different in size, the cumulative amounts of additional feed may not be in the same ratio as the ratio between dosage rates. These dosage rates and accumulated quantities must therefore be adjusted according to the requirements
of organisms in creation private individuals.[0060] To the ends of this application, the term food if refers at any food source
provided to at least one breeding body. For example, in addition to the phytoplankton and / or bacteria provided for in
Petition 870180161033, of 10/12/2018, p. 70/147
36/95 method of the present invention, at least one additional supply can be provided. This food can be of different forms and suitable ingredients to allow the growth of at least one breeding organism. For example, the feed may comprise a mixture of products of plant or animal origin in its natural, fresh or preserved state, or products derived from its industrial processing, or organic or inorganic substances, whether or not containing additives. The feed can be supplied in different forms, such as sunken tablets, extruded floating tablets, granular, crumbed, extruded smooth tablet and other shapes. An example of a fish feed is fish meal, a protein-rich food derived from processing whole fish (usually small pelagic fish and by-catches from fishing activities), as well as waste and by-products from processing facilities. of fish, such as fish waste.
[0061] Minerals and vitamins are essential for the healthy growth of breeding organisms, phytoplankton and bacteria. Minerals can comprise elements needed in trace amounts or larger amounts for healthy growth in these organisms. For example, minerals, such as zinc, calcium,
Petition 870180161033, of 10/12/2018, p. 71/147
37/95 iron, magnesium, manganese and so on are involved in certain enzymes and are essential for the maintenance of life in man, animals and plants. In some cases, vitamins facilitate the incorporation of the mineral into the enzyme, so that the activity of the enzyme is inhibited by a shortage of the mineral or vitamin. For example, zinc is involved in DNA synthesis by the zinc-containing DNA polymerase enzyme. The vitamin niacin facilitates the incorporation of zinc into the peptide subunits of the DNA polymerase enzyme. If both niacin and zinc are deficient in the body, the activity of DNA polymerase in the tissues can be reduced and the result, in both cases, is lack of growth. In aquaculture methods, where the stocking density is high, it is therefore important to supply the necessary minerals and vitamins in a form that can be efficiently taken by the body, that is, a bioavailable form. To give some non-limiting examples, bioavailable forms may include forms, such as a salt that is easily soluble in water, a floating tablet that breeding organisms can ingest, a slow-release form that is capable of releasing a constant amount of vitamins and / or minerals in the aquatic environment for consumption and assimilation by the breeding organism, phytoplankton and / or bacteria, and the like. In addition, some minerals are
Petition 870180161033, of 10/12/2018, p. 72/147
38/95 capable of providing a buffering effect on water to maintain the pH of the aqueous or aquatic environment within a certain range. For example, the mineral may comprise a reservoir of alkali metal ions that are capable of dissolving in a slightly acidic medium, such that when the pH of the aquatic environment decreases, the more alkaline metal ions dissolve in the mineral's aquatic environment. , so that the pH is kept within a certain range. The pH changes may be due to changes in the dissolved carbon dioxide resulting from photosynthesis and respiration during the day and night, respectively. Such a buffering effect can be useful in reducing the stress levels of the breeding organisms.
[0062] Therefore, in some embodiments of the present invention, the method may further comprise the step of providing at least one mineral and / or vitamin, wherein at least one of the mineral and / or vitamin is in a bioavailable form suitable for allow at least one breeding organism, phytoplankton and / or bacteria to grow. At least one of the mineral and / or vitamin can be supplied in a gradually increasing amount suitable to allow at least one breeding organism, phytoplankton and / or bacteria to grow. At least one of the mineral can be provided in an adequate amount to maintain the pH of the
Petition 870180161033, of 10/12/2018, p. 73/147
39/95 aquatic environment between about 7.5 and about 8.5. At least one of the mineral can be supplied for at least part of a predetermined period. Preferably, at least one of the mineral can be supplied in at least two predetermined periods, more preferably, over the first, second and third predetermined periods.
[0063] Beneficial and non-toxic phytoplankton can include some diatom algae and green species, which can be highly nutritious for farmed organisms. Different aquatic environments can comprise different species of green algae and diatoms according to the environment. Therefore, in some embodiments of the present invention, the phytoplankton allowed to grow may comprise at least one green alga and / or at least one species of diatom.
[0064] Some species of bacteria can be probiotic with respect to the breeding organism, which means that they confer an improvement in the health and growth of a host, which is the breeding organism. Such probiotics can be administered as a live microbial food supplement, where the breeding organism benefits by improving the balance of its intestinal microbial flora, and enzymes and vitamins produced by
Petition 870180161033, of 10/12/2018, p. 74/147
40/95 microbial supplement. These bacteria can also colonize the environment, that is, aqueous and solid phases, suppress the growth of pathogens in the environment, in addition to increasing protection against disease. They can also play an important role in the decomposition of organic matter.
Therefore, in some embodiments of the present invention, at least one of the added bacteria and / or bacteria allowed to grow can comprise at least one species of bacteria that is probiotic with respect to at least one breeding organism.
[0066] Aquaculture production changes the biochemistry of water, increasing compounds of organic and nitrogenous matter in the aquatic environment:
• Food increases organic matter and nitrogen concentration.
• Shrimps produce organic matter (feces, uneaten food, seedlings), ammonia and urea, excreted by gills and feces, respectively.
• There is an increase in natural food, phytoplankton and zooplankton, which serves as an additional food source for shrimp, but also contributes to the organic matter in the tank.
[0067] How the decomposition of organic matter in
Petition 870180161033, of 10/12/2018, p. 75/147
41/95
tank requires oxygen and releases nitrogen compounds, the demand for oxygen and the concentration of compounds nitrogenous increase both about intensity of tank. [0068] Microorganisms such as bacteria and
phytoplankton influence nutrient cycles in water and soil interphase and, indirectly, water quality parameters and stability, which are important for aquaculture production. The biochemical cycles of sulfur, silicon, phosphorus and nitrogen in the tank are strongly unbalanced when aquaculture farms operate, that is, feed intake, shrimp metabolism, etc.
[0069] Among these, the nitrogen cycle is crucial. The nitrogen cycle is very important because many organisms are involved in it and aquaculture activity changes it strongly. Too high a level of nitrogen compounds in the tank environment can be toxic to the rearing organism, for example, shrimp.
[0070] The nitrogen cycle is influenced by the biochemical activities of phytoplankton, bacteria and the breeding organism. This regulates the concentration of nitrogenous compounds in water up and down by its biological activity, as can be seen in Figure 1:
• Ammonification is the conversion of organic matter
Petition 870180161033, of 10/12/2018, p. 76/147
42/95 (uneaten food, feces, dead phytoplankton, ecdises) in ammonia (NH ₄ ), and it is carried out by heterotrophic bacteria under aerobic and anaerobic conditions. Heterotrophic bacteria (later called probiotics) break down organic matter into ammonia, decreasing biological oxygen demand and thus avoiding anaerobic conditions that induce the production of hydrogen sulfide (H ₂ S) by a different type of bacteria.
• Nitrification is the conversion of ammonia to nitrite (N0 ₂ ) and then to nitrate (NO3) by the nitrifying bacteria species Nitrosomonas and Nitrobacter, respectively, under aerobic conditions. Ammonia and nitrite are toxic at certain levels.
• The assimilation of ammonia and nitrates by phytoplankton reduces the toxicity of nitrogen compounds. The photosynthetic activity of phytoplankton reduces the concentration of CO2 in the water during the day, increasing the pH, while breathing at night produces the opposite effect. Maintaining an adequate flowering (abundance and species) of phytoplankton balances the pH and temperature in the tank water.
• Denitrification is the conversion of nitrates to atmospheric nitrogen (N ₂ ) by denitrifying bacteria.
Petition 870180161033, of 10/12/2018, p. 77/147
43/95
Atmospheric nitrogen fixed by blue-green algae. This group plays a key role in the nitrogen cycle, but they are avoided in aquaculture systems, due to their production of toxic compounds and inducing a lack of flavor in fish meat.
[0071] results from organisms.
Organic matter in an aquatic environment excretion by organisms in it and death of these
In aquaculture, the feed provided also adds organic matter to the water, either directly through uneaten food, or indirectly by increasing the excretion of farmed organisms. This is especially true when large amounts of food are added in intensive aquaculture techniques. Decomposing organic matter releases nitrogen compounds and carbon dioxide into the aquatic environment. Nitrogen compounds in particular can reach undesirable levels or concentrations that are dangerous for breeding organisms.
[0072] Phytoplankton and some species of bacteria are able to absorb nitrogen compounds and / or convert them into less toxic or non-toxic forms. However, the assimilation of ammonia or nitrate by phytoplankton is low.
Thus, in a traditional shrimp farming, very low restocking densities are preferred, to avoid
Petition 870180161033, of 10/12/2018, p. 78/147
44/95 any of the problems mentioned above. The inventors found that, surprisingly (it was not found that nitrifying bacteria maintain low ammonia and nitrates, it is common knowledge in the technical field of the inventors, but it is possible to manipulate their growth (and nitrogen cycle as well) to maintain the low concentration of nitrogen compounds), by maintaining conditions to favor their growth, nitrifying bacteria are able to keep nitrates in the aquatic environment at a low concentration to provide healthy conditions for breeding organisms.
[0073] Figure 1 shows the nitrogen cycle in an aquatic environment. Decomposing organic matter releases ammonia into water through ammonification, typically by both aerobic and anaerobic bacteria. Some naturally occurring anaerobic bacteria can be harmful and can grow to become dominant as the organic matter in the water increases. Some of these bacteria can be pathogenic for breeding organisms. However, some heterotrophic bacteria (aerobic and / or facultative anaerobes) are also able to decompose organic matter into ammonia. Although these bacteria may not be naturally present in large quantities, the inventors found that, surprisingly, by the addition of
Petition 870180161033, of 10/12/2018, p. 79/147
45/95 sufficient quantities and maintaining the conditions to favor their growth, these bacteria are able to displace the pathogenic bacteria and maintain healthy conditions for the breeding organisms. The ammonia generated by the decomposition of organic matter can then be removed by some bacteria, such as Nitrosomonas and Nitrobacter, which are, respectively, capable of converting ammonia to nitrites, and nitrites to nitrates. Nitrates can be assimilated by phytoplankton, but they can also be undesirable if they are in very large quantities, as they can promote the growth of harmful phytoplankton, such as blue-green algae. Denitrifying bacteria are therefore important for converting nitrates to non-toxic nitrogen. The inventors have found that the method of the invention in manipulating these groups of organisms during the course of aquaculture production is surprisingly effective in maintaining good water quality, even with high densities of farmed organisms. In particular, the method of the present invention manipulates groups of bacteria, such as nitrifying, denitrifying and / or heterotrophic bacteria (facultative aerobic and / or anaerobic bacteria), so that sufficient populations of such bacteria grow in parallel with the increase in organic matter and therefore supplied in
Petition 870180161033, of 10/12/2018, p. 80/147
46/95 parallel with the increase in the resulting nitrogen compounds. These populations of bacteria are then able to work in synergy to maintain concentration at a safe and convenient level for the healthy growth and development of breeding organisms. In particular,
the inventors checked what , when the present method invention is used, at bacteria nitrifying and heterotrophic (aerobic and anaerobic optional) are
able to work in synergy to reduce the concentration of nitrogen compounds.
Therefore, in some embodiments of the present invention, at least one of the phytoplankton allowed to grow, the added bacteria and / or the bacteria allowed to grow may be able to maintain the concentration of ammonia and / or nitrites and / or nitrates in the environment aquatic to a level that is non-toxic to at least one breeding organism. Preferably, at least one of the added bacteria and / or the bacteria allowed to grow may be able to maintain the concentration of ammonia and / or nitrites and / or nitrates for the aquatic environment at a level that is non-toxic to at least one organism from creation.
[0075] In some embodiments of the present invention, the bacteria grown can comprise at least one species of nitrifying bacteria.
Petition 870180161033, of 10/12/2018, p. 81/147
[0076] In some embodiments of the present invention, at least one of the added bacteria and / or the bacteria allowed to grow may comprise at least one species of denitrifying bacteria.
[0077] In some embodiments of the present invention, at least one of the added bacteria and / or the bacteria allowed to grow may comprise at least one species of aerobic and / or anaerobic facultative bacteria.
[0078] In addition, other nutrients, such as magnesium, calcium, sodium and potassium are also crucial, and intensive production can also induce mineral deficiencies in water / soil, due to the presence of limited amounts of these minerals in the tank. Absorption by the breeding organism to meet its requirements for healthy growth would thus decrease the availability of these essential minerals. The method of the present invention provides these essential minerals in a bioavailable form for the healthy growth of the organism grown in intensive aquaculture. The provision of these essential minerals also helps to maintain a stable water environment.
[0079] Therefore, in some embodiments of this specification, at least one phytoplankton nutrient provided comprises calcium, magnesium, potassium
Petition 870180161033, of 10/12/2018, p. 82/147
48/95 and sodium in forms and amounts suitable for growing phytoplankton that are non-toxic or apatogenic to at least one breeding organism.
[0080] The present invention is based on four concepts for managing organisms and their biochemical processes in the tank to balance the system:
• Phytoplankton (N: P ratio):
[0081] N: P is the relationship between the concentration of nitrogen and phosphorus in the water. Phytoplankton groups are adapted to grow with different nitrogen and phosphorus needs and, therefore, by controlling this relationship, inventors are able to control the growth of specific phytoplankton without adding any phytoplankton to the water.
[0082] By controlling the nutritional needs of phytoplankton, mainly nitrogen and phosphorus, the growth of a specific group of phytoplankton is promoted. Other important nutrients, such as calcium, magnesium, potassium, sodium, etc., are also provided.
[0083] These groups can be beneficial (green algae, diatoms) or harmful (blue green algae, dinoflagellates). The present invention balances the N: P ratio, setting it at around 16-20, promoting the growth of beneficial phytoplankton, such as green algae and diatoms.
Petition 870180161033, of 10/12/2018, p. 83/147
49/95
This N: P ratio may not be ideal for some harmful phytoplankton, such as blue-green algae, which may require a higher N: P, as seen from the approximate guide below:
Type N: P Nitrogen Fixation (Blue-Green) 42 to 125 Green ~ 30 Diatom ~ 10 Red seaweed ~ 10 Dinophyceae ~ 12
[0084] The growth of phytoplankton beneficial stabilizes the water quality (pH, temperature, compounds nitrogenous) and promotes production of food natural, highly nutritious for shrimp. • Minerals: [0085] The present invention provides Forms of
essential minerals and ions absorbable (bioavailable) in the water of the fish / shrimp tank to absorb directly from the water through the gills, fins and other membranes. This will alleviate mineral deficiencies in the tank and shrimp, and will balance the acid-bases in the water.
• Probiotics (C: N ratio):
[0086] Bacteria use nitrogen, organic matter, as an energy source, thus reducing its concentration in water and acting as a bioremediator. To ensure continuous bioremediation, an external source of
Petition 870180161033, of 10/12/2018, p. 84/147
50/95 carbon (molasses) is added to facilitate bacterial growth and performance. The amount of carbon source to be added is calculated according to the nitrogen concentration of the water and guarantees an adequate C: N ratio.
[0087] In the present invention, high performance probiotics (heterotrophic bacteria) are provided to decompose organic matter in the water column and bottom tank, and nutrients and / or micronutrients are provided to promote their growth. Aerobic and facultative anaerobic bacteria and micronutrients that effectively degrade organic matter at the bottom of the tank, even under low oxygen conditions.
[0088] Generally, the activity bacterian, reduces it toxicity in compounds nitrogenous and avoids conditions anaerobic what induce production of sulfide hydrogen
toxic. It helps to recover aerobic conditions after the death of phytoplankton (dead phytoplankton is organic matter that requires oxygen for their degradation), increases nitrification by reducing toxic nitrogen compounds and promotes a clean bottom environment that allows for larger benthic organisms.
[0089] Addition of the continuous probiotic suppresses the outbreak of pathogenic bacteria by competing their growth in the water column, deep tank and digestive tract of the shrimp.
Petition 870180161033, of 10/12/2018, p. 85/147
51/95 • ORP:
[0090] Nitrifying bacteria require aerobic conditions and essential nutrients to grow and develop. The present invention provides the nutrients for nitrifying bacteria and facilitates a system with high ORP (Oxidation Reduction Potential) between 100 and 350 mV. A high ORP value is related to oxidative, aerobic conditions that favor nitrification, degradation of organic matter and biological removal of phosphorus.
[0091] Promoted aerobic conditions prevent the formation of hydrogen sulfide and undesirable fermentation at the bottom of the tank.
[0092] This invention also provides the nutrients essentials that promote the growth in nitrifying bacteria. [0093] This invention regulate the reactions
biochemicals mentioned above and organisms involved to minimize the toxic effects derived from the concentration of organic matter in the water.
[0094] Nutrient supply ratios, such as the ratio of nitrogen to phosphorus (N: P) or carbon to nitrogen (C: N) are used in the method of the present invention to determine the amount of nutrients
Petition 870180161033, of 10/12/2018, p. 86/147
52/95 to supply, thus controlling the growth of phytoplankton and bacteria. These proportions refer to the desired atomic relative abundance of these elements and are understood by aquaculture technicians. As an example, the concentrations of phosphorus and nitrogen can be calculated with conventional chemical methods, including all compounds containing phosphorus and nitrogen present in a water sample collected in the aquatic environment. This is then compared to the desired N: P ratio and the appropriate amount of phytoplankton nutrients containing nitrogen and / or phosphorus can be provided. Likewise, the carbon concentration needed to achieve a given C: N ratio is calculated by multiplying the nitrogen concentration with the C: N ratio, and the mass of the carbon source needed to achieve the carbon concentration is calculated from the aquatic used for aquaculture, volume of the environment quantity of carbon present bioavailable in the used carbon source. Carbon is typically used to promote the growth of bacteria, so the carbon source must provide carbon-containing compounds that are bioavailable for bacteria. Suitable carbon sources can be any non-toxic carbon-containing energy source, such as molasses and brown sugar, but not limited to them.
Petition 870180161033, of 10/12/2018, p. 87/147
53/95 aquaculture technician would be able to adjust the amount of carbon source used according to the concentration of carbon available in the carbon source unit.
[0095] Different proportions of N: P can promote the growth of different groups of phytoplankton and, similarly, different proportions of C: N can promote the growth of different groups of bacteria. The inventors have found that the method of the present invention is surprisingly effective in promoting the growth of the desired groups of phytoplankton and bacteria by maintaining certain ranges of N: P and C: N ratios. N: P values as low as 10 induce the growth of harmful phytoplankton. Excessively high or low values induce the growth of harmful phytoplankton and the present invention balances this around 16 to 20.
[0096] In particular, in some embodiments of the present invention at least one phytoplankton nutrient can be provided in an amount suitable to maintain an N: P ratio in the aquatic environment of between about 16 to about 20. Preferably, the N ratio : P can be maintained in this range for at least a predetermined period. More preferably, the N: P ratio can be maintained in this interval between the first, second and third predetermined periods.
Petition 870180161033, of 10/12/2018, p. 88/147
[0097] In some embodiments of the present invention, at least one bacterial nutrient can be provided in an amount suitable for maintaining a C: N ratio in the aquatic environment suitable for growing bacteria, which is non-toxic to at least a creative organism. In some embodiments, at least one nutrient for the bacteria can be provided in an adequate amount to maintain a proportion of C: N in the aquatic environment suitable for growing the bacteria, which is capable of maintaining the concentration of ammonia and / or nitrites and / or nitrates in the aquatic environment at a level that is non-toxic to at least one of the rearing organism. According to any of these embodiments, the C: N ratio can be between about 6 and about 10. Preferably, the C: N ratio can be maintained in this range over at least a predetermined period. More preferably, the C: N ratio can be maintained in this interval between the first, second and third predetermined periods. According to any of these embodiments, at least one nutrient for the supplied bacteria can comprise at least one carbon source. Suitable carbon sources can be any non-toxic carbon source, such as molasses or brown sugar, but not limited to them.
Petition 870180161033, of 10/12/2018, p. 89/147
55/95 [0098] In some embodiments of the present invention, at least one bacterial nutrient is provided in an amount adequate to maintain an Oxidation Reduction Potential (ORP) in the aquatic environment between about + 100 mV to about + 350mV . The inventors have found that this can help to promote the growth of the desired bacteria. In particular, maintaining the ORP in this range can increase nitrification, reduce the oxygen demand needed for the degradation of organic matter and suppress the production of hydrogen sulfide.
[0099] In addition to the bacteria nutrients above, the desired groups of bacteria may require more nutrients in trace amounts for healthy growth. Therefore, in some embodiments of the present invention, at least one nutrient for bacteria can comprise micronutrients in forms and amounts suitable for growing bacteria that are non-toxic or apatogenic to at least one breeding organism. In some embodiments, the provided bacterial nutrient comprises micronutrients in forms and amounts suitable for growing bacteria that are capable of maintaining the concentration of ammonia and / or nitrites and / or nitrates for the aquatic environment at a level that is non-toxic to at least one of the creative organism.
Petition 870180161033, of 10/12/2018, p. 90/147
56/95 [0100] In some embodiments of the present invention, the aquatic environment may comprise the aqueous phase and may also comprise the solid phase in a tank and may further comprise any layer of organic matter and / or any cavity in fluid communication with the aqueous phase.
[0101] The present invention can comprise three phases that can correspond to the first, second and third predetermined periods. For example, some embodiments of the present invention can comprise the following steps:
• Phytoplankton phase: (also referred to as the first predetermined period)
Production phase: for example, Culture Day (DOC) 1 to DOC 35-40.
Organisms: photoautotrophic organisms (phytoplankton) are predominant.
Water quality: in the early stages of production there is a low content of organic matter, since the feed rates are not very high (shrimp of small size and thus low amounts of feed are added to the tank), having mainly inorganic importance .
[0102] Phytoplankton, which rely on inorganic matter to grow, are abundant due to water fertilization (supplemented with nitrogen, phosphorus and others
Petition 870180161033, of 10/12/2018, p. 91/147
57/95 essential nutrients) and allows the growth of natural food. This natural food consists of zooplankton and other highly nutritious benthic organisms that provide additional food for shrimp, which is very important in the initial production stage. The balanced phytoplankton population promotes a more stable environment and, therefore, water quality (pH, temperature, oxygen).
• Phytoplankton and probiotic phase: (also referred to as the second predetermined period)
Production phase: for example, DOC 35-40 to DOC 70-75.
Organisms: decrease in photoautotrophic organisms (phytoplankton) and increase in chemoautotrophic (nitrifying bacteria) and heterotrophic bacteria.
Water quality: increase in the organic matter content is proportional to the feeding rates that increase in the tank. Shrimp, larger in size, require more food to grow and, therefore, produce more metabolic residues (feces, humus, etc.) that contribute to the increase in feeding rates, build the organic matter content of the water. The aqueous environment, in turn, requires higher rates of degradation of organic matter to ammonia by heterotrophic bacteria and even more from nitrification to nitrites and nitrates by nitrifying bacteria. At this stage, there is still an abundance of phytoplankton, but less important than in
Petition 870180161033, of 10/12/2018, p. 92/147
58/95 previous phase.
• Probiotic Phase: (also referred to as the third predetermined period)
Production phase: for example, DOC 70-75 for the harvest.
Organisms: chemoautotrophic bacteria (nitrifying bacteria) and heterotrophic bacteria are predominant.
Water quality: very high loads of organic matter (food, feces, humus, etc.) are decomposed by heterotrophic bacteria, while nitrifying bacteria convert the resulting ammonia into inorganic nitrogen (nitrites and nitrates).
[0103] At previous phases can also be represented at following table that ranks organisms by power supply:
Power supply Autotrophic Autotrophic and Heterotrophic Autotrophic and Heterotrophic DOC 1 to 35-40 35-40 70-75 for the harvest Phase Phytoplankton Phytoplankton and probiotic Phytoplankton and probiotic Organisms Phytoplankton, Bacterium Nitrifying and dominant zooplankton nitrifying and heterotrophic, phytoplankton (less) heterotrophic Note: Abundant natural food and low organic matter, mainly INORGANIC Matterincreased organic (food), ammonia and urea. High levels of ORGANIC matter, ammonia and urea.
Petition 870180161033, of 10/12/2018, p. 93/147
59/95 [0104] An autotrophic is an organism that produces complex organic compounds from simple inorganic molecules, using light energy (phytoplankton), or inorganic chemical reactions (nitrifying bacteria). Some other examples include nitrifying bacteria such as nitrosomonas Nitrosomonas spp. and nitrobacter Nitrobacter spp., denitrifying bacteria and blue-green algae.
[0105] A heterotrophic is an organism that cannot fix carbon and uses sources of organic carbon. Some examples are heterotrophic or probiotic bacteria, zooplankton and farmed organisms such as shrimp.
[0106] Some organisms are defined as myxotrophic because they can act as autotrophic and heterotrophic. Examples include some species of phytoplankton and zooplankton.
[0107]
The present invention is described as a mixotrophic system because the system uses and manipulates both autotrophic (phytoplankton and nitrifying bacteria) and heterotrophic (bacteria) organisms throughout the production cycle.
[0108]
The production cycle, from restocking to harvest, induces a succession of abundance of inorganic compost to organic compost in the water, which,
Petition 870180161033, of 10/12/2018, p. 94/147
60/95, in fact, is parallel to the predominance of autotrophic bacteria (phytoplankton and nitrifying bacteria) to heterotrophic (probiotic) throughout the above mentioned phases.
[0109] Surprisingly, the inventors have found that it is possible to manipulate groups of autotrophic and heterotrophic organisms to grow in synergy according to the method of the invention, allowing the productivity of an aquaculture tank to be safely increased.
[0110] The present invention provides essential nutrients to promote the growth of phytoplankton, natural food and bacterial growth, stabilize water quality and ensure compliance with the nutritional requirements of shrimp (nutrients provided and growth of natural food provided). The present invention also supplies beneficial bacteria (also known as probiotics) to the tank, which break down organic matter and create a clean environment. This activity of the organisms balances the biological parameters (natural food, presence of pathogens), physical (temperature) and chemical (oxygen, pH, organic matter, nitrogen, etc.) of the water, allowing a larger and safer aquaculture production.
[0111] Phytoplankton, probiotic bacteria, their biochemistry and the benefits of applying each one
Petition 870180161033, of 10/12/2018, p. 95/147
61/95 of them for aquaculture are known to aquaculture producers and the scientific community.
However, the method of the present invention provides an original protocol for managing these factors in synergy, providing energy nutrients to manage phytoplankton, bacteria and ambient water. The present invention promotes highly nutritious natural food (zooplankton, polychaetes, etc.) and suppresses the production of outbreaks of pathogens through the control of water, soil and microflora in the breeding organism, by and fish. The invention thus achieves, for example, shrimp the surprising benefits, such as:
Have an ecologically balanced system.
Minimize fluctuations in water and soil quality.
- Reduce stress for the breeding organism.
- Increase the ideal load capacity.
- Increase production, improving the growth rate, FCR (Feed Conversion Rate), the survival rate and daily growth of the breeding organism.
- Reduce the energy cost due to the management of aeration and zero water exchange.
- Reduce compounding costs for food and
Petition 870180161033, of 10/12/2018, p. 96/147
62/95 chemicals (such as lime, iodine).
- Manage the soil and sediment of the tank.
- Improve the quality of water and effluents and minimize the environmental impact.
- Minimize the risk of bankruptcy due to illness or low production.
- Long-term sustainability of production.
- All at an economical cost and easy to manage.
[0112] The present invention balances water quality to avoid stress, disease outbreaks or slow growth and thus increase productivity, health, growth and survival. All of these benefits allow you to safely increase production and reduce food and energy costs.
[0113] Thus, it can be seen that over the three predetermined periods that constitute a production cycle, there may be an increase in the concentration of organic matter in the aquatic environment, especially as increasing amounts of food can be added. At the beginning of production, in the first predetermined period, the matter in the aquatic environment can be essentially inorganic. At the end of the third predetermined period of time, matter in the aquatic environment can be
Petition 870180161033, of 10/12/2018, p. 97/147
63/95 predominantly organic in nature. This change from inorganic to organic matter may correspond to the passage from the predominance of phytoplankton to bacteria from the first predetermined period to the third predetermined period. Along with these changes, there may also be a change in the aquatic environment for substantially chemoautotrophic and heterotrophic organisms.
[0114] Therefore, in some embodiments of the present invention, in the third predetermined period of time at least one breeding organism, phytoplankton and bacteria present in the aquatic environment are substantially chemoautotrophic and heterotrophic.
[0115] As the method of the present invention uses and manipulates autotrophic (phytoplankton and nitrifying bacteria) and heterotrophic (bacteria) organisms throughout the production cycle to increase production volume and quality, the method of the present invention is known as a mixotrophic aquaculture method or a mixotrophic system. Therefore, for the purposes of this specification, the method of the present invention can be referred to interchangeably as a mixotrophic aquaculture method or a mixotrophic system.
[0116] The method of the present invention is capable of reducing the aeration needs of the production of
Petition 870180161033, of 10/12/2018, p. 98/147
64/95 aquaculture. For the purposes of the present invention, aeration refers mainly to the enrichment of the aquatic environment with oxygen and can also more generally comprise the promotion of gas exchange between the gas and the aquatic environment. For example, paddle aerators are sometimes used in the aquaculture tank to promote the gas exchange between the tank water and the atmosphere in the process of removing carbon dioxide from the water and enriching the water with oxygen.
[0117] An aerator refers to any device used to aerate the aquatic environment and can be one or more than one unit and types of aeration device suitable for use alone or in combination in aquaculture. For example, the aerator can operate by diffusion or hydraulic action. A hydraulic aerator can comprise, for example, a cascade, a sprinkler, an ejector or an air inlet head connected to a pump through a pipe, or it can comprise a surface aerator, such as a simple open impeller or a pump centrifuge, placed on or near the surface of the aquatic environment to mix water with the atmosphere. A diffusion-type aeration apparatus may comprise, for example, a root-type fan, a fan, a compressor or a membrane pump to pump air into the aquatic environment through
Petition 870180161033, of 10/12/2018, p. 99/147
65/95 of a porous material, such as a perforated tube.
[0118] Examples of aerators include paddle aerators, cyclonic aerators, dissolved oxygen conditioners, Venturi oxygenators, source aerators, air injectors, tube aerators, long arm aerators and circular aerators and cyclonic reducers are examples of aerators that are available commercially. Oxygen injection and pure oxygen diffusion systems are also increasingly used as an aeration device. These can be more expensive and can sometimes be reserved for use as emergency aerators to quickly alleviate hypoxia conditions. Aerators can be used alone or in combination with other aerators to satisfy the demand for oxygen from organisms in the aquatic environment.
[0119] However, aerators can be energy intensive and / or expensive. Aquaculture production is often limited by the amount of aeration used to aerate the aquatic environment. For simplicity, the amount of aeration can be compared approximately at different farms in terms of the number of horsepower (hp) of aerators installed to aerate the aquatic environment and the number of hours that aerators are used per day. Alternatively, aerator-based equivalents
Petition 870180161033, of 10/12/2018, p. 100/147
66/95 standards, such as a paddle aerator, can be calculated by comparing the effective increase in the oxygen content of the water.
[0120] The method of the present invention can also eliminate the need for water exchange in aquaculture. Water exchange refers to a common practice in aquaculture for draining or discharging water from the aquatic environment and replacing the water discharged with better quality water, thus improving the water quality of the aquatic environment. For example, in some aquaculture tanks next to a water source, such as a river, poor quality water (eg, with low dissolved oxygen content, high carbon dioxide content and / or high concentration of compost nitrogen) can be discharged from the tank into the river downstream of the tank and fresh water upstream of the tank can be supplied to the tank, thereby improving the quality of the water in the tank (higher dissolved oxygen content, concentrations carbon dioxide and nitrogen compounds). This cycle of flushing and replacing water, known as water exchange, can also help flush out excess levels of phytoplankton to reduce nutrient concentrations and to regulate salinity. It is an intense energy and can result in pollution of the natural water source with poor water.
Petition 870180161033, of 10/12/2018, p. 101/147
67/95 quality (with high content of carbon dioxide and nitrogen compounds, and / or low content of oxygen). The method of the present invention does not comprise water exchange where water exchange is not necessary to maintain or improve the quality of the water exchange. More specifically, the method may not necessarily comprise the exchange of water from the aquatic environment for at least one of the first, second and / or third predetermined periods. However, normal water inputs to the aquatic environment can be imposed by evaporation and / or excessive infiltration of water from the tank through the walls of the tank, which can in some cases be water permeable. This replacement of lost water may be necessary, even with the method of the present invention.
[0121] Therefore, in some embodiments of the present invention, the method may not comprise discharging water from the aquatic environment for at least one of the first, second and / or third predetermined periods. Preferably, the method may not comprise discharging water from the aquatic environment during any of the first, second and / or third predetermined periods.
[0122] The embodiments of the present invention refer essentially to an aquaculture method of at least one farmed organism, in which the farmed organism
Petition 870180161033, of 10/12/2018, p. 102/147
68/95 is not phytoplankton or bacteria. At least one of the breeding organisms can be selected from the group consisting of fish, crustaceans, molluscs, algae and / or invertebrates. For example, at least one of the breeding organisms can be selected from the group consisting of tilapia, catfish, milk fish, groupers, perchs, carp, snakeheads, catlas, sturgeons, eels, mullets, rohus, sea bass , sparids, rabbit fish, prawns, river prawns, crabs, lobsters, crayfish, oysters, shellfish, mussels, scallops, clams, tame ears, sea cucumbers, sea urchins. In particular, at least one breeding organism can be fish and / or shrimp.
[0123] In particular, the system of the present invention is particularly well suited for raising fish and / or shrimp. Thus, much of the remaining description can be directed towards embodiments where the breeding organism is fish and / or shrimp. It must be understood, however, that the system is also well suited for the breeding of other aquatic farmed organisms.
[0124] Aquaculture production can proceed through a production cycle, which can have a desired production cycle duration, depending on the desired end products of aquaculture. For example, the length of the desired production cycle may vary according to the species of
Petition 870180161033, of 10/12/2018, p. 103/147
69/95 breeding organisms and their growth requirements. The production cycle can begin with the repopulation of the aquatic environment with young farmed organisms, for example, with larval or post-larval shrimp. The production cycle can end the harvest of the grown organisms. However, grown breeding organisms are not necessarily fully mature. The desired length of the production cycle can also vary depending on the desired maturity of the breeding organisms at the time of harvest.
[0125] In some embodiments, the method of the present invention can further comprise a step of determining a desired duration of the production cycle for at least one breeding organism and the first predetermined period can be from about 30% to about 50 % of the intended production cycle duration and may start with the restocking of the aquatic environment; the second predetermined period can be from about 30% to about 50% of the desired duration of the production cycle duration and can start with the end of the first predetermined period and end with the beginning of the third predetermined period of time; and the third predetermined period of time can be from about 0% to about 40% of the desired duration of the production cycle and can start with the end of the second
Petition 870180161033, of 10/12/2018, p. 104/147
70/95 predetermined period of time and end with the harvest of at least one breeding organism.
[0126] In particular embodiments, the first predetermined period can be from about 30% to about 40% of the desired duration of the production cycle and can start with the restocking of the aquatic environment; the second predetermined period can be from about 30% to about 40% of the desired duration of the production cycle, starting with the end of the first predetermined period of time and ending with the beginning of the third predetermined period of time; and the third predetermined period of time can be from about 30% to about 40% of the desired duration of the production cycle, starting with the end of the second predetermined period of time and ending with the harvest of at least one breeding organism. In some preferred embodiments, the three predetermined periods can be of equal duration, that is, each one being a third of the production cycle.
[0127] In some embodiments, the method of the present invention can be directed to an aquaculture method of at least one farmed organism, in which at least one of the farmed organism comprises shrimp, the first predetermined period may be between about 35 to about 40 days, the second predetermined period of time can be between about 35 to about 40
Petition 870180161033, of 10/12/2018, p. 105/147
71/95 days and the third predetermined period in the desired production cycle can be at least for about 5 days and can end with the harvest of at least one breeding organism.
[0128] For the purposes of this application, load capacity refers to the amount (expressed in weight or number) of organism creation that a given aquatic environment is capable of supporting. The load capacity is limited by a factor, which, on the farm, is usually oxygen, then ammonia and carbon dioxide.
[0129] Restocking density refers to the weight or number of breeding organisms carried out per unit area or volume. Restocking densities depend on the breeding organism and its tolerance to the stress of greater agglomeration.
[0130] Thus, restocking, for the purposes of this invention, refers to the introduction of one or more organisms into the aquatic environment in the work of the invention. For example, the method of the invention may comprise an initial step of adding larvae or post-larvae of the organism grown in the aquatic environment, which may already have phytoplankton and bacteria.
[0131] The restocking of the aquatic environment may be at a restocking density for a number of young farms per unit volume or area
Petition 870180161033, of 10/12/2018, p. 106/147
72/95 of the aquatic environment. The area unit can be in accordance with the water surface area. The present invention allows for a surprisingly high restocking density. For example, some embodiments of the present invention may be directed to an aquaculture method of at least one farmed organism, including shrimp, and may comprise a restocking step of at least about 200 shrimp per square meter of the aquatic environment at the beginning of the first predetermined period. In particular, some embodiments may comprise restocking at least about 300 shrimp per square meter of the aquatic environment at the beginning of the first predetermined period of time.
[0132] The rate in feed conversion or ‘FCR if refers to the ratio between O dry weight of foods for the organisms creation to allow them grow and the gain weight per part From organisms creation after O
growth. FCR is a measure of the efficiency of converting food to fish - for example, an FCR of 2.8 means that 2.8 kg of food is needed to produce 1 kg of live weight of fish. Different species of culture organisms have different FCRs, depending on the aquaculture method used. For example, tilapia can have a typical FCR of 1.6 to 1.8. Typical shrimp can have an FCR
Petition 870180161033, of 10/12/2018, p. 107/147
73/95 above about 1.5. As can be seen from the data provided below, the inventors surprisingly found that, when using the method of the present invention, an average of FCR 1.29 is obtained per shrimp culture, compared to an average of FCR 1 , 59 when using traditional methods. This means that the increase in mass for the reared shrimp from restocking for harvest was on average 0.775 times the mass of the cumulative feed provided, compared to only 0.629 times when using traditional methods. However, the present invention is not limited to embodiments, such as these. The same effect of improving FCR applies to all breeding bodies. Values may vary, but there is always a clear improvement.
[0133] Therefore, in some embodiments of the present invention, the use of the method of the present invention to allow breeding organisms to grow can increase the mass of the breeding organisms by at least about 0.7 times the mass of at least additional food
provided to leave of start of first period predetermined for O Final of third period predetermined •[0134] With techniques of traditional breeding,
restocking density is limited by the above problems
Petition 870180161033, of 10/12/2018, p. 108/147 mentioned. For example, in shrimp farming it is impossible to reach a restocking density of 300 to 400 post larvae (PL) per square meter of the farm's surface in a sustainable manner, since a high restocking density would induce an unstable system subject to failure and disease. .
[0135] Surprisingly, the inventors have found that the use of the method of the present invention to manipulate the aquatic environment (water and soil quality management) allows for a much higher stocking density in aquaculture. The inventors manipulated the activities of phytoplankton and bacteria to balance the system and increase production safely.
[0136] As can be seen from the data below in improving production performance, comparing traditional aquaculture methods and the method of the present invention, the benefits of using the present invention include: increased stocking density, increased growth rate, increased production, reduced FCR, reduced (or improved) aeration costs, etc., all in a sustainable manner.
[0137] In addition, the present invention is unique and inventive in providing a complete protocol adapted to any species, or breeding peculiarities to manage
Petition 870180161033, of 10/12/2018, p. 109/147
75/95 the tank environment throughout the crop cycle to increase production rates, minimizing disease outbreaks at much higher densities than in general aquaculture practice, and at reduced costs.
[0138] For example, in major shrimp-producing countries, such as Thailand, India or Ecuador, previous methods achieved a restocking density of 200, 100 and 30 post-larvae per square meter, respectively. With the present invention, more than 200 and typically about 300 to about 400 post-larvae per square meter can be supplied.
[0139] Traditional methods of aquaculture breeding may comprise aquatic environments with populations of phytoplankton and bacteria. However, there is no protocol to manipulate the activity of phytoplankton and bacteria populations. So an aquaculturist
qualified not I would expect to control these organisms, phytoplankton and bacteria, their biochemistry and thus
manipulate them to improve the quality and volume of aquaculture production. This description provides a new and inventive method of aquaculture comprising a protocol that can manipulate these populations of organisms by regulating the energy and nutrients that these organisms obtain from water.
Petition 870180161033, of 10/12/2018, p. 110/147
76/95 [0140] In another aspect of the present invention, an aquaculture system capable of carrying out the method according to any aspect of the present invention, the system comprising:
(a) an aquatic environment comprising at least one breeding organism, phytoplankton and bacteria and / or means to provide such an environment;
(b) at least one phytoplankton nutrient that provides a means to provide at least one phytoplankton nutrient to the aquatic environment;
(c) at least one phytoplankton nutrient detection means to detect at least one concentration of phytoplankton nutrients in the aquatic environment;
(d) at least one means of supplying bacterial nutrient to supply at least one bacterial nutrient to the aquatic environment;
(and) fur any less one middle in addition of bacterium p > ara add fur any less an bacterium to the environment aquatic; and (f) fur any less one middle in detection nutrient in bacteria for to detect fur minus one concentration in nutrient of bacteria at the environment aquatic. [0141] For to provide The amount proper in
nutrients and / or bacteria and / or to maintain nutrients and / or bacteria at a given concentration, a system
Petition 870180161033, of 10/12/2018, p. 111/147
77/95 aquaculture can be used, which can comprise various detection means operatively coupled to various supply means. For example, detection or testing devices for determining the concentration of substances and / or organisms in the water, which can indicate when more nutrients and / or bacteria should be supplied in the water. The system can be manual, automated or partially automated, for example, it can comprise an automatic nutrient dispensing device and / or automated sampling systems and detection systems, such as those available at http://www.aquacultureequipment.co.uk and / or http://www.campbellsci.com.au/products and / or http://www.ysi.com/products.php. However, the invention is not limited to such embodiments and includes embodiments in which all or part of the characteristics of the aquaculture system depend on human operators.
[0142] In some embodiments of the present invention, the aquaculture system may further comprise:
(g) at least one phytoplankton nutrient maintenance means operatively coupled to at least one phytoplankton nutrient supply means and / or at least one phytoplankton nutrient detection means to maintain phytoplankton nutrients in a concentration
Petition 870180161033, of 10/12/2018, p. 112/147
78/95 suitable for growing phytoplankton; and (h) at least one means of maintaining the concentration of bacterial nutrients operatively coupled to at least one means of supplying bacterial nutrients and / or at least one means of detecting bacterial nutrients to maintain bacterial nutrients at a adequate concentration to grow bacteria, in which phytoplankton and bacteria are allowed to grow in a proportion of phytoplankton: bacteria greater than 1 during the first predetermined period;
phytoplankton and bacteria are allowed to grow in a second proportion of phytoplankton: predetermined bacteria during the second predetermined period of time, where the second proportion of phytoplankton: predetermined bacteria is less than the first proportion of phytoplankton: predetermined bacteria; and phytoplankton and bacteria are allowed to grow in a third proportion of phytoplankton: predetermined bacteria during the third predetermined period of time, where the third proportion of phytoplankton: predetermined bacteria is less than the second proportion of phytoplankton: predetermined bacteria.
[0143] There may be some cases where growth
Petition 870180161033, of 10/12/2018, p. 113/147
Excessive 79/95 of even the desired phytoplankton and bacteria can induce harmful situations derived from low oxygen. Therefore, in some embodiments of the present invention, the aquaculture system can further comprise:
(i) at least one phytoplankton detection means to detect the concentration of phytoplankton grown;
(j) at least one bacterial detection means for detecting the concentration of bacteria allowed to grow;
(k) at least one means of maintaining the concentration of phytoplankton nutrients operatively coupled with at least one means of supplying phytoplankton nutrients and / or at least one means of detecting phytoplankton to prevent a new disposition of phytoplankton nutrients when concentration of the phytoplankton phytoplankton allowed to grow reaches a first predetermined concentration, until the concentration of phytoplankton allowed to grow falls below the first predetermined concentration; and (1) at least one means of maintaining the concentration of nutrients less one means of bacteria operatively coupled to the supply of nutrient bacteria and / or at least one means of detecting bacteria, to avoid
Petition 870180161033, of 10/12/2018, p. 114/147
80/95 additional provision of bacterial nutrients and / or additional addition of bacteria when the concentration of bacteria allowed to grow reaches a second predetermined concentration, until the concentration of bacteria allowed to grow falls below the second predetermined concentration. The first and second predetermined concentrations may indicate the overgrowth of phytoplankton and bacteria, respectively.
[0144] Bacteria detection means and phytoplankton detection means may comprise the manual investigation of water samples in the laboratory and / or apparatus that is capable of counting and / or identifying bacteria and / or phytoplankton. For example, means of detecting bacteria may comprise a genetic analysis device, such as that provided for at http://www.springerlink.com/content/v5443m2823833888/. However, in most cases, the use of such devices may not currently be possible on a large scale due to cost reasons.
[0145] In common aquaculture practice, populations of phytoplankton and bacteria may not be measured directly by the apparatus to count the concentrations of phytoplankton and bacteria. Instead, indirect means can be used to indicate excessive growth of
Petition 870180161033, of 10/12/2018, p. 115/147
81/95 phytoplankton and / or bacteria. For example, in some embodiments of the present invention, the aquaculture system may further comprise an apparatus for obtaining a Secchi disk visibility reading for the aquatic environment and the nutrient supply means may provide no additional phytoplankton nutrients when the The visibility of the Secchi disk in the aquatic environment is less than about 30 cm, and can return to supply phytoplankton nutrients when the visibility of the Secchi disk in the aquatic environment increases to above about 30 cm.
[0146] The downs levels of oxygen dissolved at the environment aquatic also can signal the need in stop the supply more nutrients from bacteria. Per
therefore, in some embodiments of the present invention, at least one bacterial detection means can comprise an apparatus for measuring dissolved oxygen in the aquatic environment and the means of supplying bacterial nutrients can provide no additional bacterial nutrients when dissolved oxygen in the aquatic environment it is less than about 3.5 mg / L, and can continue to provide nutrients from bacteria when the oxygen dissolved in the aquatic environment increases to above about 3.5 mg / L. Likewise, other environmental parameters may indicate
Petition 870180161033, of 10/12/2018, p. 116/147
82/95 the need to stop, increase or decrease amounts of food and / or phytoplankton and / or bacteria nutrients
provided. Sensors suitable forparameters are, for example,http://www.ysi.com/products.php, to detectfound thesein http://www.aquacultureequipment.co.ukand / or http://www.campbellsci.com-.au/products. Example 1Performance data records of production it's from
water quality [0147] The following table summarizes the difference in aquaculture production performance between a traditional shrimp farming system and a shrimp farm using the method of the present invention.
Table 1.
Traditional Myxotrophic Improvement (%) Restocking density (PL / sqm) 85 209 146 Average Body Weight(g) 13.62 14.34 5 ADGR (g / day) 0.16 0.20 26 Survival(%) 65, 6 81.6 24 FCR 1.59 1.29 19 kg collected / hp 266 677.68 155 kg food per day / hp 4.90 12.29 151
Petition 870180161033, of 10/12/2018, p. 117/147
83/95 * hp = horsepower installed in the tank.
[0148] The more detailed data tables follow after the discussion of the results below.
• The increase in stocking density allows the harvesting of a larger volume of shrimp without affecting the survival rate. The data show that there is even an improvement in the survival rate and a larger volume of shrimp can be collected by the installed horsepower aerator (= energy saving) when the method of the present invention is implemented.
• Aeration management is improved so that greater volume of shrimp can be produced with the same aeration in a safe manner, or the amount of installed aeration (hp) can be reduced to produce the same volume and thus the costs of energy are reduced.
• Higher feed volume (kg) can be provided with the method of the present invention by installed horsepower. This means that the system is balanced and more organic matter is allowed in the system without affecting the health of the shrimp, because maintaining good water quality is being maintained. The average body weight is also greater in less days of culture.
• There is a decrease in FCR and an increase in the average daily growth rate (g / day), because the quality of the water is not slowing the growth of the shrimp. The shrimp
Petition 870180161033, of 10/12/2018, p. 118/147
84/95
healthy will metabolize food improve, therefore, will grow a lot better, improving The rate in conversion (FCR).[0149] A quality management gives water and of soil is a
important part of the creation, therefore, water quality data is also shown to provide an insight into the management of the system.
TANKS E5, E6, E7 and E8 (traditional shrimp farming system) • pH is a logarithmic function which means that an increase or decrease of the unit is a ten-fold change. Therefore, the unstable pH (as shown in the data for the traditional shrimp farm) during the production cycle leads to shrimp stress, therefore, to decreased productivity, and this can induce the disease outbreak.
• Ammonia spikes, as shown in the data, cause shrimp stress, reducing productivity and leading to disease outbreaks.
• Increase in organic matter should be gradual. The sudden increases, as in tanks E5 and E6, are due to the unbalanced system, where organic matter is accumulated and this requires high oxygen demand. Anaerobic zones can be created, leading to the formation of toxic gas with all
Petition 870180161033, of 10/12/2018, p. 119/147
85/95 the harmful health effects of shrimp.
TANKS Dl, D6, D10, D4 and D5 (mixotropic shrimp farming system) • The stable pH is a sign of balanced phytoplankton activity and acid-base concentration.
• The data shows how ammonia is always kept low due to continuous nitrification and ammonia removal from phytoplankton. The nitrite gradually increases in parallel with the increase in organic matter from restocking to harvest. Ammonia levels close to zero do not mean that there is no ammonia, but this is due to the method of measuring ammonia, which provides a result of ammonia close to zero (in the tables, also ammonia values of zero do not mean that ammonia, but there is a low concentration). Ammonia is required all the time for nitrifying bacteria to grow and convert it to nitrite and nitrate.
• Decreased visibility of the Secchi disk is related to the flowering of phytoplankton at the beginning of repopulation. Then it slowly stabilizes, while the organic matter accumulates slightly due to the increase in the volume of food.
[0150] The following tables provide more detailed data
Petition 870180161033, of 10/12/2018, p. 120/147
86/95 detailed on aquaculture production performance (Total Production (= kg harvested), FCR, Survival Rate (SR,%), daily food vs. installed horsepower and total amount of food supplied vs. horsepower installed) and water quality in a traditional shrimp farming system, and on a shrimp farm using the method of the present invention.
Table 2 - Shrimp farming (traditional), DATA AND PRODUCTION INDICATORS Tank Date fueled Tank size (sqm) DOC Density (PL / sqm) Total fueled (pcs) Average Body Weight(g) ADGR (g / day) Survival (%) E5 6-Feb-11 4700 83 85 399000 15.33 0.18 52.4 E6 4-Feb-11 4000 88 84 336600 15.95 0.18 78.0 E7 6-Feb-11 4100 87 85 346500 15.98 0.18 57.0 E8 3-Feb-11 4000 81 86 342700 7.21 0.09 75.0 Average8513.62 0.16 65.6
Tank Date fueled FCR Total food (kg) Average daily feed (kg) Harvest biomass (kg) Horsepower (hp) installed kg collected / hp kg food day / hp E5 6-Feb-11 1.79 5737.0 69.1 3205.0 14 229 4.94 E6 4-Feb-11 1.35 5651.7 64.2 4186.5 11 381 5.84 E7 6-Feb-11 1.77 5585.4 64.2 3155.6 11 287 5.84 E8 3-Feb-11 1.44 2669.1 33.0 1853.6 11 169 3.00 Average1.59266 4.90
Table 3 - Blue Aqua Mixotrophic System for Intensive Shrimp Farming PRODUCTION DATA AND INDICATORS Tank Date fueled Tank size (sqm) DOC Density (PL / sqm) Total fueled (pcs) Average Body Weight (g) ADGR (g / day) Survival (%) D1 11-Dec-n 3400 71 214 727800 14.21 0.20 95.1 D2 11-Dec-11 3900 72 202 788800 14.14 0.20 89.0 D6 11-Dec-11 5300 74 206 1089400 14.83 0.20 87.3 D9 11-Dec-11 4300 69 196 841900 13.45 0.19 72.2
Petition 870180161033, of 10/12/2018, p. 121/147
87/95
D10 11-Dec-11 3700 70 208 771300 15.53 0.22 84.6 D7 14-Dec-11 5000 71 194 971000 15.12 0.21 67.7 D4 16-Dec-11 5900 72 222 1310000 14.22 0.20 81.5 D3 17-Dec-11 5100 71 220 1124400 14.24 0.20 64.6 D5 17-Dec-11 5000 70 208 1038800 14.20 0.20 78.0 D8 22-Dec-11 4700 71 217 1020200 13.46 0.19 96.2 Average20914.34 0.20 81.6
Table 4 - Blue Aqua Mixotrophic System for Intensive Shrimp Farming PRODUCTION DATA AND INDICATORS Tank Date fueled Tank size (sqm) DOC FCR Total food (kg) Average food / day (kg) Biomass Collected (kg) Horsepower installed (hp) kg collected / hp installed Daily food (kg) / hp D1 11-Dec-11 3400 71 1.25 12300.0 173.2 9837.3 16 615 10.83 D2 11-Dec-11 3900 72 1.24 12345.3 171.5 9922.5 16 620 10.72 D6 11-Dec-11 5300 74 1.23 17326.0 234.1 14109.2 20 705 11.71 D9 11-Dec-11 4300 69 1.37 11197.0 162.3 8172.3 12 681 13.52 D10 11-Dec-11 3700 70 1.20 12135.0 173.4 10139.1 16 634 10.83 D7 14-Dec-11 5000 71 1.40 13935.0 196.3 9937.8 15 663 13.08 D4 16-Dec-11 5900 72 1.29 19534.0 271.3 15179.2 20 759 13.57 D3 17-Dec-11 5100 71 1.46 15163.0 213.6 10351.2 16 647 13.35 D5 17-Dec-11 5000 70 1.32 15142.0 216.3 11512.9 16 720 13.52 D8 22-Dec-11 4700 71 1.14 15016.0 211.5 13206.0 18 734 11.75 Average1.29677.68 12.29
Petition 870180161033, of 10/12/2018, p. 122/147
Table 5 - Shrimp farming (traditional technique), WATER QUALITY
'sF co co O O 35.9 'sF co o> r * - 37.9 LO ^ r LOCOr— r * -O co F * - CM LO Or * -LO CO co r * - o> LOr * - r— r * - O O ^ r F * - co O Or * - co O O 'sF r * - r * - O'sF CM CO LO LO co O o> r * - O CM cm O coO LO O co LOcm r— r * - O O co r * - co O O co r * - r * - O O CO r * - r * - O'sF LO co r * - CO CM co o>Oco O> co co o> LOO co o> LO co r * - O O CM co r * - O O CM co r * - O O CO co r * - O O CO LO co LO LO LO r * - co * "O CT> co co LO O i <T co * "O o> co * "O CT> LO co O O CM LO co O O CM LO co O O CM LO co O O CM LO r * - LO ^ r LO r * - LO 'sF o> o> LO O CT> o> coO co o> o>O co o> CMO LO ^ r r * - O O CM ^ r co O O CM 'sF r * - O O CM 'sF co O O CM LO CM LO LO LO 'sF CO CM coOCM * "O O CM CMO co CM CM co ^ r co O O CM ^ r co O O CM 'sF co O O CM 'sF co O O CM LO r * - LO ^ r LO co LOLO coOLO o>O LO LO OLO O CO co co O Oco r * - O O co CO co O O CM CO CO O O CM LO o> LO CM LO co LO CO co coO r * - co coO cm CO CMO co CO O>O CT> CM co O OCM co O O CM CM co O OCM r * - O Oco LO ^ r LO o> LO CO^ r* " r * -CMO cocoO CT>CMO CT> CM co O OCM co O O CM CM co O OCM co O O LO ^ r LO co LO 'sF LO LO ^ r CMO co ^ r CMO r * - 'sF r * -O LO 'sF O ^ fco O O co O O r * - O O CMco O O CM LO CM LO LO O> LO COr * -O coo>O LO* "O coo>O r * - r * - r * - O O ^ r r * - r * - O O LO r * - co O Or * - r * - O Oσ) 0) 0) 0) co CO CO co O O O O τc <CB τc <CB τc <CB τc <CB "Co *dd) ppm org; "Co *(PP t Q_ Q_ org: "Co *dd) ppm org: "Co *dd) ppm org: ©COCO 0coCO 0COCO 0COCO O O O OO"O<d O"O<d O"O<d O"O<d <-> X F M— » co <-> X F M— » co <-> X F M— » co <-> X F M— » co O Q_ < zO Q_ < zO Q_ < zO Q_ < zLO co r * - co LU LU LU LU CD CD CD CD 0 0 0 0 σ σ σ σ ç ç ç ç co co co co 1- 1- 1- 1-
Petition 870180161033, of 10/12/2018, p. 123/147
89/95
Table 6.
DOC DiscSecchi(cm) pH Ammonia(ppm) Nitrite(ppm) Organic matter(g) 7 100 7.8 0 0.1 77.1 14 100 7.9 0.5 0.1 61.9 21 85 8 0 0.05 98.6 28 70 7.9 0 0.05 90.1 35 55 7.9 0 0.2 99.9 42 45 7.8 0 0.2 111.2 49 45 7.6 0 1 107.4 56 40 7.8 0.5 1.8 108.7 63 35 7.7 1 1 106.2 70 30 7.6 2 3 102.3
Tank D6
DOC DiscSecchi(cm) pH Ammonia(ppm) Nitrite(ppm) Organic matter(g) 14 100 8.2 0 0.05 108.7 21 90 8 0 0.05 68.2 28 75 7.9 0 0.05 97.3 35 60 7.9 0 0.05 106.2 42 40 7.7 0 0.05 108.7 49 35 7.8 0 0.4 99.9 56 30 7.8 0 1 108.7 63 25 7.7 0 6 113.8 70 30 7.6 0 6 111.2
Petition 870180161033, of 10/12/2018, p. 124/147
90/95
DOC DiscSecchi(cm) pH Ammonia(ppm) Nitrite(ppm) Organic matter(g) 14 100 8.1 0 0.05 106.2 21 75 8.2 0 0 64.5 28 50 7.9 0 0.05 109.9 35 35 8 0 0.05 99.9 42 30 7.8 0 0.0549 30 7.8 0 0.4 106.2 56 30 7.8 0 163 30 7.7 0 3 112.5 70 30 7.7 0 10 109.9 77 30 7.7 0 10 86
DOC DiscSecchi(cm) pH Ammonia(ppm) Nitrite(ppm) Organic matter(g) 7 100 7.9 0 0.05 60.7 14 100 8.2 0 0.05 101.1 21 85 8.1 0 0.05 54.4 28 65 7.8 0 0.05 96.1 35 40 7.8 0 0.1 104.9 42 30 7.8 0 0.2 111.2 49 30 7.8 0 0.6 103.6 56 30 7.7 0 1 108.7
Petition 870180161033, of 10/12/2018, p. 125/147
91/95
63 30 7, 7 0 6 107.4 70 25 7, 7 0 6 109.9 77 25 7, 6 0 10 96.1
Tank D5
DOC DiscSecchi(cm) pH Ammonia(ppm) Nitrite(ppm) Organic matter(g) 7 100 7.9 0 0.05 60.7 14 100 8.1 0 0.05 65.7 21 100 8 0 0.05 79.6 28 85 7.8 0 0.05 72 35 70 7.8 0 0.05 87.2 42 50 7.8 0 0.05 79.6 49 30 7.7 0 0.05 96.1 56 30 7.6 0 0.2 88.5 63 30 7.6 0 1 89.7 70 30 7.7 0 6 102.4
Example 2
Economic analysis of a 3.1 MT shrimp production on a farm using a conventional shrimp farming method compared to an equivalent farm using the (mixotrophic) method of the present invention
GENERAL INFORMATION • Production volume equal to 3.1 MT.
• Feed costs equal to 1.24 SGD / kg.
Petition 870180161033, of 10/12/2018, p. 126/147
92/95 • FCR = 1.59 vs. 1.29 (Conventional vs Mixotrophic).
• 85 days of culture (DOC).
• Total tank area equal to 1.6 ha.
LIMING INFORMATION:
• Application 3x / week at 200 kg / ha.
• Lime cost is 0.124 SGD / kg (Mixotrophic system does not use lime).
[0151] Traditional aquaculture methods include a step of applying various neutralizing compounds of calcium or calcium and magnesium acids to the aquaculture tank, for example, before filling with water. This is known as liming and has three important advantages: 1) Liming can increase the fertilization effect. 2) Liming helps to avoid large fluctuations in pH. 3) Liming also adds calcium and magnesium, which are important in the proper development of a breeding organism. Materials such as agricultural limestone, slag, slaked lime, quicklime and liquid lime have been used for lime tanks. Liming materials may comprise one or more of carbonate, hydrogen carbonate, hydroxide, and calcium and magnesium oxide salts. However, as can be seen from the data above, several applications are generally necessary and can be expensive. The present invention
Petition 870180161033, of 10/12/2018, p. 127/147
93/95 allows the reduction of operating costs in the aquaculture tank by balancing the pH and manipulating the environment in such a way that liming is not necessary.
ENERGY COSTS INFORMATION:
• Energy cost 0.124 SGD / kWh.
Aeration:
• Operating hours / day = 8 • Operating days = 85 • Engine efficiency 80% in both cases.
• Conventional system: 12 hp installed (266 kg / hp, from the performance table)) • Mixotrophic system: 5 hp installed (677 kg / hp; from the performance table)).
Pumping • 2 hp pump with 80% efficiency in both cases.
• Hours of operation / day = 4 (Conventional System).
• Hours / day operation = 0.5 (Mixotrophic System,
Zero Water Exchange, only replenishment of evaporation).
[0152] Aeration and water exchange also contribute significantly to energy costs for aquaculture production, especially in aquaculture sites where energy costs are high. The method of the present invention
Petition 870180161033, of 10/12/2018, p. 128/147
94/95 manipulates the environment in such a way that there are very low seration requirements for a given amount of production. The data above show the improvement in performance (expressed as volume of shrimp (kg) harvested by horsepower installed and volume of food diary (kg) per installed power) when a traditional shrimp farm adopts the method of the present invention. From these data it can be seen that using the same horsepower installed as the aerators, the production volume can be increased three times, from 266 to 677 kg per horsepower installed.
[0153] Water exchange is also not necessary in the process of the present invention, although water evaporated from the aquatic environment must be replenished. In addition, energy and operating cost savings are therefore due to the fact that there is no need to operate water pumps to drain or discharge water from aquaculture tanks, and having to refill drained aquaculture tanks before proceeding. with the next stage of aquaculture.
[0154] The production costs for the respective farms are compared in Figure 2, showing that the reduction of costs in feeding, liming, aeration and water / pumping water changes leads to a total saving of approximately SGD 3232 to 3.1 MT of shrimp.
Petition 870180161033, of 10/12/2018, p. 129/147
95/95 [0155] The matter claimed here is not limited to embodiments that solve the drawbacks, or that operate only in environments such as those described above. Instead, this foundation is provided only to illustrate the area of exemplary technology where some of the embodiments described here can be practiced. The bibliographic references cited in this specification are listed for convenience in the form of a list of references and added at the end of the examples. All content of such bibliographic references is incorporated by reference.

权利要求:
Claims (35)
[1]
1. Aquaculture method of at least one breeding organism characterized by the fact that it comprises the steps:
(i) providing an aquatic environment comprising at least one breeding organism, phytoplankton and bacteria;
(ii) provide at least one phytoplankton nutrient and at least one bacterial nutrient during a first predetermined period, allowing phytoplankton and bacteria to grow at a first predetermined proportion of phytoplankton: bacteria greater than 1, preferably at least about 60 : 40;
(iii) providing at least one phytoplankton nutrient and at least one bacterial nutrient during a second predetermined period, allowing phytoplankton and bacteria to grow in a second predetermined proportion of phytoplankton: bacteria, where the second predetermined proportion of phytoplankton: bacteria is less than the first predetermined proportion of phytoplankton: bacteria, preferably between about 75:25 to about 25:75; and (iv) providing at least one phytoplankton nutrient and at least one bacterial nutrient during a third predetermined period, allowing phytoplankton and bacteria to grow in a third predetermined proportion
Petition 870180161033, of 10/12/2018, p. 131/147
[2]
2/16 phytoplankton: bacteria, where the third predetermined proportion of phytoplankton: bacteria is less than the second predetermined proportion of phytoplankton: bacteria, preferably less than about 40:60;
where the first predetermined period is from about 30% to about 50% of the desired duration of the production cycle and begins with the restocking of the aquatic environment;
the second predetermined period is about 30% to about 50% of the desired duration of the production cycle and begins with the end of the first predetermined period and ends with the beginning of the third predetermined period; and the third predetermined period is from about 0% to about 40% of the desired duration of the production cycle and begins with the end of the second predetermined period and ends with the harvest of at least one of the rearing organism.
2. Method according to claim 1, characterized in that at least one of the phytoplankton nutrient and at least one of the bacterial nutrient are provided during the first, second and third predetermined periods in the respective concentrations suitable for growing phytoplankton and bacteria in the first, second and third predetermined proportions of phytoplankton:
Petition 870180161033, of 10/12/2018, p. 132/147
[3]
3/16 bacteria.
Method according to any one of claims 1 to 2, characterized in that at least one phytoplankton nutrient is provided during the first, second and third predetermined periods in decreasing concentrations suitable for growing phytoplankton and bacteria in the first, second and third predetermined proportions of phytoplankton: bacteria.
[4]
Method according to any one of claims 1 to 3, characterized in that at least one bacterial nutrient is provided during the first, second and third predetermined periods in decreasing concentrations suitable for growing phytoplankton and bacteria in the first, second and third predetermined proportions of phytoplankton: bacteria.
[5]
Method according to any one of claims 1 to 4, characterized by the fact that it further comprises the addition of bacteria to the aquatic environment, in which the added bacteria are able to maintain the concentration of ammonia and / or nitrites and / or nitrates in the aquatic environment at a level that is non-toxic to at least one breeding organism and / or where the bacteria are non-toxic or apatogenic to at least one of the breeding organism, and at least one of the added bacteria
Petition 870180161033, of 10/12/2018, p. 133/147
4/16 can comprise a species of denitrifying bacteria; or a kind of optional aerobic and / or anaerobic bacteria.
[6]
Method according to claim 5, characterized in that the bacterium is added during the first, second and third predetermined periods in increasing concentrations suitable to allow phytoplankton and bacteria to grow in the first, second and third predetermined proportions of phytoplankton :bacteria.
[7]
Method according to any one of claims 1 to 6, characterized in that the first predetermined proportion of phytoplankton: bacteria is at least 75:25.
[8]
Method according to any one of claims 1 to 7, characterized in that the first predetermined proportion of phytoplankton: bacteria is at least about 90:10.
[9]
Method according to any one of claims 1 to 8, characterized in that the second predetermined proportion of phytoplankton: bacteria is between at least about 60:40 to about 40:60.
[10]
10. Method according to any one of claims 1 to 9, characterized by the fact that:
Petition 870180161033, of 10/12/2018, p. 134/147
5/16 the aquatic environment has a visibility of the Secchi disk between about 60 cm to about 30 cm during the first predetermined period;
the aquatic environment has a visibility of the Secchi disk between about 40 cm to about 20 cm during the second predetermined period; and the aquatic environment has a visibility of the Secchi disk between about 70 cm to about 60 cm during the third predetermined period.
[11]
Method according to any one of claims 1 to 10, characterized in that it further comprises the step of providing at least one additional feed for at least one breeding organism to grow, the additional feed being provided in a proportion of 1: A: B in the first, second and third predetermined periods, respectively, where A is between about 3 to about 15 and / or B is between about 0 to about 30, where the additional feed may comprise a mixture of products of plant or animal origin in their natural, fresh or preserved state, or products derived from their industrial processing, or organic or inorganic substances, whether or not containing additives.
[12]
12. Method according to claim 11, characterized by the fact that A is between about 5 to
Petition 870180161033, of 10/12/2018, p. 135/147
6/16 about 10 and / or B is between about 15 to about 20.
[13]
13. Method according to any one of claims 1 to 12, characterized in that it further comprises the step of supplying at least one mineral and / or vitamin, wherein at least one of the mineral and / or vitamin is in a bioavailable form suitable to allow at least one breeding organism, phytoplankton and / or bacteria to grow, in which the mineral can be selected from magnesium, calcium, sodium and potassium; and where the bioavailable form may include forms such as a salt that is easily soluble in water, a floating tablet that the breeding organisms can ingest, a slow release form that is capable of releasing a constant amount of vitamins and / or minerals in the aquatic environment for consumption and assimilation by the breeding organism, phytoplankton and / or bacteria.
[14]
14. Method according to claim 13, characterized by the fact that at least one mineral and / or vitamin is supplied in a gradually increasing amount suitable to allow at least one breeding organism, phytoplankton and / or bacteria to grow .
[15]
15. Method according to claim 13 or 14, characterized by the fact that at least one mineral is
Petition 870180161033, of 10/12/2018, p. 136/147
7/16 supplied in an adequate amount to maintain the pH of the aquatic environment between about 7.5 and about 8.5.
[16]
16. Method according to any one of claims 1 to 15, characterized in that at least one of the phytoplankton nutrient provided comprises calcium, magnesium, potassium and sodium in forms and amounts suitable for growing phytoplankton that are non-toxic or apatogenic to at least one of the breeding organism.
[17]
17. Method according to any one of claims 1 to 16, characterized in that at least one of the phytoplankton nutrient is supplied in an amount adequate to maintain an N: P ratio in the aquatic environment between about 16 to about 20.
[18]
18. Method according to any one of claims 1 to 17, characterized in that at least one of the bacterial nutrient is supplied in an amount suitable for maintaining a C: N ratio in the aquatic environment suitable for growing bacteria that is non-toxic to at least one of the breeding organism, where the said C: N ratio is between about 6 to about 10.
[19]
19. Method according to any one of claims 1 to 18, characterized in that at least one of the bacterial nutrient is supplied in a
Petition 870180161033, of 10/12/2018, p. 137/147
8/16 adequate amount for maintaining a C: N ratio in the aquatic environment suitable for growing bacteria that is capable of maintaining the concentration of ammonia and / or nitrites and / or nitrates in the aquatic environment at a level that is non-toxic to at least minus one of the rearing organism, where the said C: N ratio is between about 6 to about 10.
[20]
20. Method according to any of claims 18 and 19, characterized in that at least one bacterial nutrient supplied comprises at least one carbon source, which can comprise any non-toxic carbon-containing energy source, such as molasses and Brown sugar.
[21]
21. Method according to any one of claims 1 to 20, characterized in that at least one bacterial nutrient is provided in an amount adequate to maintain an Oxidation Reduction Potential (ORP) in the aquatic environment between about +100 mV up to about + 350mV.
[22]
22. Method according to any one of claims 1 to 21, characterized in that at least one bacterial nutrient supplied comprises micronutrients in forms and amounts suitable for growing bacteria that are non-toxic or apathogenic to
Petition 870180161033, of 10/12/2018, p. 138/147
9/16 at least one of the breeding body.
23. Method, according to any of the claims 1 to 22, characterized by the fact that the
The bacterial nutrient provided comprises micronutrients in forms and amounts suitable for growing bacteria that are capable of maintaining the concentration of ammonia and / or nitrites and / or nitrates in the aquatic environment at a level that is non-toxic to at least one of the rearing organism.
24. Method, according to any of the claims 1 to 23, characterized by the fact that the
The aquatic environment comprises the aqueous phase and the solid phase in a tank, as well as any layer of organic matter and / or any cavity in fluid communication with the aqueous phase.
25. Method, according to any of the claims 1 to 24, characterized by the fact that in the
third predetermined period at least one of the rearing organism, phytoplankton and bacteria present in the environment
aquatic are substantially chemoautotrophic and
heterotrophic.
26. Method, according to any of the claims 1 to 25, characterized by the fact that the method excludes step of discharging water from the environment aquatic during at least one of the first, second and / or
Petition 870180161033, of 10/12/2018, p. 139/147
10/16 third predetermined periods.
[23]
27. Method according to any one of claims 1 to 26, characterized in that the method excludes the step of discharging water from the aquatic environment during any one of the first, second and / or third predetermined periods.
[24]
28. Method according to any one of claims 1 to 27, characterized in that at least one of the breeding organism is different from phytoplankton or bacteria.
[25]
29. Method according to any one of claims 1 to 28, characterized by the fact that at least one of the rearing organism is selected from the group consisting of fish, crustaceans, molluscs, algae and / or invertebrates.
[26]
30. Method according to any one of claims 1 to 29, characterized by the fact that at least one of the breeding organism is selected from the group consisting of tilapia, catfish, milk fish, groupers, perch, carps, snakeheads, catlas, sturgeons, eels, mullets, rohus, sea bass, spiders, rabbit fish, prawns, river prawns, crabs, lobsters, crayfish, oysters, shellfish, mussels, scallops, clams, ears- sea cucumbers and sea urchins.
Petition 870180161033, of 10/12/2018, p. 140/147
11/16
[27]
31. Method according to any one of claims 1 to 30, characterized by the fact that at least one of the breeding organism is fish and / or shrimp.
[28]
32. Method according to any one of claims 1 to 31, characterized in that it further comprises a step of determining a desired duration of the production cycle for at least one of the rearing organism, and in which:
the first predetermined period is about 30% to about 50% of the desired duration of the production cycle and begins with the restocking of the aquatic environment;
the second predetermined period is about 30% to about 50% of the desired duration of the production cycle and begins with the end of the first predetermined period and ends with the beginning of the third predetermined period; and the third predetermined period is from about 0% to about 40% of the desired duration of the production cycle and begins with the end of the second predetermined period and ends with the harvesting of at least one from the rearing organism.
[29]
33. Method according to any of claims 1 to 32, characterized by the fact that:
the first predetermined period is from about 30% to about 40% of the desired duration of the production cycle and
Petition 870180161033, of 10/12/2018, p. 141/147
12/16 begins with restocking the aquatic environment;
the second predetermined period is from about 30% to about 40% of the desired duration of the production cycle duration and begins with the end of the first predetermined period and ends with the beginning of the third predetermined period; and the third predetermined period is from about 30% to about 40% of the desired duration of the production cycle and
begins with the end of second predetermined period and ends with the harvest of hair least one of the organism in creation • 34 Method, of wake up with any of the
claims 1 to 32, characterized by the fact that at least one breeding organism comprises shrimp, the first predetermined period is between about 35 to about 40 days, the second predetermined period is between about 35 to about 40 days and the The third predetermined period in the desired production cycle is at least about 5 days and ends with the harvest of at least one of the rearing organism.
[30]
35. Method according to any one of claims 1 to 34, characterized in that at least one of the rearing organism comprises shrimp and the method comprises restocking at least about 200
Petition 870180161033, of 10/12/2018, p. 142/147
13/16 shrimp per square meter of the aquatic environment at the beginning of the first predetermined period.
[31]
36. Method according to any one of claims 1 to 35, characterized in that at least one rearing organism comprises shrimp and the method comprises restocking at least about 300 shrimp per square meter of the aquatic environment at the beginning of the first predetermined period of time.
[32]
37. Aquaculture system capable of carrying out the method as defined in any one of claims 1 to 36, characterized by the fact that it comprises:
(a) an aquatic environment comprising at least one breeding organism, phytoplankton and bacteria, and / or means for providing such an environment;
(b) at least one phytoplankton nutrient providing a means to supply at least one phytoplankton nutrient to the aquatic environment;
(c) at least one phytoplankton nutrient detection means to detect at least one concentration of phytoplankton nutrients in the aquatic environment;
(d) at least one bacterial nutrient supply means for supplying at least one bacterial nutrient to the aquatic environment;
(e) at least one means of adding bacteria to
Petition 870180161033, of 10/12/2018, p. 143/147
14/16 add at least one bacterium to the aquatic environment;
(f) at least one bacterial nutrient detection means to detect at least one bacterial nutrient concentration in the aquatic environment;
(g) at least one phytoplankton nutrient maintenance means operatively coupled to at least one phytoplankton nutrient supply means and / or at least one phytoplankton nutrient detection means; and (h) at least one means of maintaining the bacterial nutrient concentration operatively coupled with at least one means of supplying bacterial nutrient and / or at least one means of detecting bacterial nutrient, wherein:
phytoplankton and bacteria are allowed to grow in a predetermined proportion of phytoplankton: bacteria from:
more than 1 during the first predetermined period;
phytoplankton and bacteria are allowed to grow in a second predetermined proportion of phytoplankton: bacteria during the second predetermined period, where the second predetermined proportion of phytoplankton: bacteria is less than the first predetermined proportion of phytoplankton: bacteria; and phytoplankton and bacteria are allowed to grow in a third predetermined proportion of
Petition 870180161033, of 10/12/2018, p. 144/147
15/16 phytoplankton: bacteria during the third predetermined period, where the third predetermined proportion of phytoplankton: bacteria is lower than the second predetermined proportion of phytoplankton: bacteria.
[33]
38. Aquaculture system, according to claim 37, characterized by the fact that it also comprises:
(i) at least one phytoplankton detection means to detect the concentration of phytoplankton grown;
(j) at least one bacterial detection means for detecting the concentration of bacteria allowed to grow;
(k) at least one means of maintaining the phytoplankton nutrient concentration operably coupled to at least one phytoplankton nutrient supply means and / or at least one phytoplankton detection means; and (l) at least one means of maintaining the bacterial nutrient concentration operatively coupled to at least one bacterial nutrient supply means and / or at least one bacterial detection means.
[34]
39. Aquaculture system according to claim 38, characterized by the fact that at least one of the phytoplankton detection means comprises a
Petition 870180161033, of 10/12/2018, p. 145/147
16/16 apparatus for obtaining a Secchi disk visibility reading for the aquatic environment and in which the phytoplankton nutrient supply means provide no additional phytoplankton nutrients when the Secchi disk visibility from the aquatic environment is less than about 30 cm, and resumes the supply of phytoplankton nutrients when the visibility of the Secchi disk in the aquatic environment increases to more than about 30 cm.
[35]
40. Aquaculture system according to claim 38, characterized in that at least one means of detecting bacteria comprises an apparatus for measuring dissolved oxygen in the aquatic environment and in which the means of supplying nutrient bacteria provide no additional bacterial nutrients when oxygen dissolved in the aquatic environment is less than about 3.5 mg / L, and resumes the bacterial nutrient supply when oxygen dissolved in the aquatic environment increases to more than about 3.5 mg / L L.

类似技术:

---

公开号 | 公开日 | 专利标题

---

US20190008126A1|2019-01-10|Mixotrophic method of aquaculture

---

Arnold et al.2009|High-intensity, zero water-exchange production of juvenile tiger shrimp, Penaeus monodon: an evaluation of artificial substrates and stocking density

---

CN104430095B|2017-01-11|Fresh water farming method of penaeus vannamei boone

---

JP6267810B2|2018-01-24|Oyster farming method

---

US20190045756A1|2019-02-14|Dark-bright integrated greenhouse system in intensive recirculating eco-aquaculture and aquaculture method

---

CN100372510C|2008-03-05|Artificial culture of shrimp parents

---

Levy et al.2017|Marine periphyton biofilters in mariculture effluents: nutrient uptake and biomass development

---

KR101489660B1|2015-02-04|METHOD FOR CULTUERING CATADROMOUS FISH| BY USING RECIRCULATING AQUACULTUER SYSTEM AND SEAWATER ACCLIMATION

---

Tremblay-Gratton et al.2018|Bioremediation efficiency of Palmaria palmata and Ulva lactuca for use in a fully recirculated cold-seawater naturalistic exhibit: effect of high NO3 and PO4 concentrations and temperature on growth and nutrient uptake

---

CN105638525A|2016-06-08|Closed type circulating water breeding method of hexagrammos otakii

---

Datta2012|Management of water quality in intensive aquaculture

---

Venkateswarlu et al.2019|A study on water quality parameters in shrimp L. vannamei semi-intensive grow out culture farms in coastal districts of Andhra Pradesh, India

---

Tidwell et al.1997|Growth, survival, and biochemical composition of freshwater prawns Macrobrachium rosenbergii fed natural food organisms under controlled conditions

---

Kawamoto1961|The influence of excretory substances of fishes on their own growth

---

CN107087560A|2017-08-25|Penaeus Vannmei and the foundation and its application of golden pomfret ecological polyculturing pattern

---

CN102100190B|2013-07-17|Method for cultivating epinephelus septemfasciatus offspring seeds

---

Sallenave2012|Understanding water quality parameters to better manage your pond

---

Guttman et al.2019|An integrated Ulva-periphyton biofilter for mariculture effluents: Multiple nitrogen removal kinetics

---

CN105454144A|2016-04-06|Pelteobogrus fulvidraco fish culturing water body eutrophication early warning method and application of same

---

CN105340804A|2016-02-24|Yellow catfish ecological symbiotic cultivation method

---

CN105052837A|2015-11-18|Breeding method of larval stage feed of coregonus peled fry

---

CN108849645A|2018-11-23|A kind of freshwater fish culturing method

---

KR101317143B1|2013-10-15|Incubation method for a food organism of a sea bass species.

---

Hartman et al.2015|Carp aquaculture in Europe and Asia

---

Osinga et al.2008|Feeding corals in captivity: uptake of four Artemia-based feeds by Galaxea fascicularis

同族专利:

---

公开号 | 公开日

---

MX2014015745A|2015-06-05|

---

IN2014DN09072A|2015-05-22|

---

MX358353B|2018-08-15|

---

SG11201407585QA|2014-12-30|

---

US9992979B2|2018-06-12|

---

CN104582475B|2018-04-20|

---

US20150296752A1|2015-10-22|

---

WO2013191642A1|2013-12-27|

---

US20190008126A1|2019-01-10|

---

CN104582475A|2015-04-29|

---

BR112014031658A2|2017-06-27|

引用文献:

---

公开号 | 申请日 | 公开日 | 申请人 | 专利标题

---

CN109997744A|2018-08-16|2019-07-12|三门县绿洋特种水产养殖专业合作社|Feed formula of the autumn crab in aquaculture net cage and fat up method|

---

CN110292009A|2019-07-22|2019-10-01|广西壮族自治区水产科学研究院|The litopenaeus vannamei ecological breeding method regulated and controled based on biological feed and water quality biological|US4137869A|1977-03-28|1979-02-06|Kipping Vernon L|System and method for production of marine food using submerged platform|

---

US5353745A|1992-09-29|1994-10-11|Fahs Ii Richard W|Aquaculture system and methods for using the same|

---

GB9503127D0|1995-02-09|1995-04-12|Peroxythai Ltd|Improvements or modifications to a process for farming crustacea|

---

JP2772772B2|1995-03-09|1998-07-09|中央水産研究所長|Production method of seaweed detritus|

---

US6327996B1|2000-01-07|2001-12-11|The Oceanic Institute|Biosecure zero-exchange system for maturation and growout of marine animals|

---

US7482151B2|2000-05-19|2009-01-27|Aquaria, Inc.|Method of using ammonia-oxidizing bacteria|

---

EP1302103A1|2000-07-06|2003-04-16|Nagase &amp; Co., Ltd.|Method of culturing zooplankton, apparatus for culturing by the same, and culture obtained by the same|

---

US7544501B2|2003-10-09|2009-06-09|Aquaria, Inc.|Nitrite-oxidizing bacteria and methods of using and detecting the same|

---

AU2005206778A1|2004-01-09|2005-08-04|Richard S. Brauman|Method and system for aquaculture production|

---

CN101264980A|2008-04-23|2008-09-17|中国水产科学研究院南海水产研究所|Sea water culture environment multiple state position biological restoring method|

---

WO2010102202A2|2009-03-06|2010-09-10|Kenneth William Gregg|Composition for controlling fish|

---

CN101885554A|2009-05-12|2010-11-17|上海楚水水产科技有限公司|Method for treating aquaculture wastewater|

---

EP2459725B1|2009-07-27|2019-01-16|The University Of Wyoming Research Corporation|Biological clean fuel processing systems and methods|

---

CN101869086B|2010-07-09|2013-11-27|浙江省淡水水产研究所|Environmentally-friendly culture method for black carps|

---

US20120285392A1|2011-05-13|2012-11-15|LiveFuels, Inc.|Deep water nutrient recovery system|WO2015179788A1|2014-05-23|2015-11-26|Nch Corporation|Method for improving quality of aquaculture pond water|

---

US10766799B2|2014-05-23|2020-09-08|Nch Corporation|Method for improving quality of aquaculture pond water using a nutrient germinant composition and spore incubation method|

---

CN104782548A|2015-04-24|2015-07-22|西双版纳飞龙渔业有限责任公司|Hemibagrus nemurus pond high-yield breeding method|

---

CN105409848A|2015-12-10|2016-03-23|广东海洋大学|Method for controlling bacterial diseases by co-culturing Siganus ConcatenaluJ and Penaeus japonicus|

---

CN106259080B|2016-08-09|2019-06-25|中国水产科学研究院南海水产研究所|It is a kind of that water cultural method is changed based on the Penaeus Vannmei high zero density for nitrifying cenobium regulation water quality|

---

US20200002665A1|2016-12-14|2020-01-02|Heliae Development, Llc|Methods of Applying Ammonia Toxicity and Inducing Nitrogen Uptake in Microalgae Cultures|

---

CN106577412A|2017-01-09|2017-04-26|嵇学范|Artificial breeding method for stone snails|

---

CN107094679A|2017-06-20|2017-08-29|盐城工学院|A kind of method that application chlorella cultivates river crab seedling|

---

WO2019040787A1|2017-08-24|2019-02-28|L&B Patent Inc.|System and method for integrated multi-trophic aquaculture|

---

CN108142326A|2017-11-17|2018-06-12|浙江师范大学|The artificial domestication method of She Minnow|

---

CN108541628A|2018-03-20|2018-09-18|中国水产科学研究院珠江水产研究所|A kind of method for culturing seedlings of the hybridized snakehead fish based on pond|

---

CN108496857A|2018-04-19|2018-09-07|广州普麟生物制品有限公司|A kind of closed zero-emission batch production method for breeding tilapia|

---

CN109006634B|2018-07-31|2020-05-29|中国水产科学研究院渔业机械仪器研究所|Circulating water prawn water color regulation and control three-dimensional culture system and control method|

---

CN109362621A|2018-10-31|2019-02-22|四川东雨农业开发有限公司|The high-quality cultural method of rice field shrimp|

---

CN110050728A|2019-04-30|2019-07-26|云南省渔业科学研究院|A kind of sturgeon cultivation method in plateau waters|

---

CN110521648A|2019-10-15|2019-12-03|中国水产科学研究院黄海水产研究所|A kind of joint nitrification-assimilation prawn circulating water cultivation method|

---

CN110800661A|2019-11-22|2020-02-18|烟台市海洋经济研究院|Ecological prevention and control method for gastric ulcer disease of auricularia juveniles of stichopus japonicus|

---

CN110839569A|2019-12-16|2020-02-28|江苏省农业科学院宿迁农科所|Method for cultivating wild rice field eel seedlings in spring|

---

CN110973022A|2019-12-31|2020-04-10|海南晨海水产有限公司|Efficient and high-yield grouper and penaeus vannamei boone polyculture method|

---

WO2021221745A1|2020-05-01|2021-11-04|Atlantic Sapphire Ip, Llc|Method for detection and mitigation of compound producing microbiota in an aquaculture system|

---

CN111436391B|2020-04-21|2022-03-04|宁波大学|Domestication method for enabling scylla paramamosain fries to adapt to saline-alkali soil water area culture|

---

CN111406684A|2020-05-06|2020-07-14|陈晓方|Breeding method for preventing May plague by low-density rotation catching and rotation releasing of crayfish|

---

CN112005937A|2020-09-04|2020-12-01|泗阳县双高水产科技有限公司|Ecological breeding mode of weever and corbicula fluminea|

---

CN112293305A|2020-10-30|2021-02-02|广东大麟洋海洋生物有限公司|Method for three-dimensional mixed culture of egg-shaped pompano and scallop|

---

CN112673997A|2021-01-07|2021-04-20|湄潭县永兴米业有限公司|Rice field control and circulating breeding method|

法律状态:
2018-03-27| B15K| Others concerning applications: alteration of classification|Ipc: A01K 61/00 (2017.01), A01K 67/00 (2006.01) |

---

2018-04-03| B06F| Objections, documents and/or translations needed after an examination request according art. 34 industrial property law|

---

2018-09-11| B07A| Technical examination (opinion): publication of technical examination (opinion)|

---

2019-02-05| B09A| Decision: intention to grant|

---

2019-03-26| B16A| Patent or certificate of addition of invention granted|Free format text: PRAZO DE VALIDADE: 20 (VINTE) ANOS CONTADOS A PARTIR DE 18/06/2012, OBSERVADAS AS CONDICOES LEGAIS. |

优先权:

---

申请号 | 申请日 | 专利标题

---

PCT/SG2012/000216|WO2013191642A1|2012-06-18|2012-06-18|Mixotrophic method of aquaculture|

---