阅读说明：本技术 处理水的组合物和方法 (Compositions and methods for treating water ) 是由 P.比扎 P.琼斯 D.卡明斯 A.莱昂斯 C.布思比 M.德塞利 于 2018-03-16 设计创作，主要内容包括：描述了处理水以促进水生生物健康的方法。方法可包括将经粉碎高岭土引入水中。经粉碎高岭土可具有一定粒径分布,使得至少30%重量的颗粒通过60目筛。例如,经粉碎高岭土可以为具有小于300μm的d<Sub>50</Sub>直径的干燥粉末的形式。经粉碎高岭土可在没有分散剂和/或表面活性剂的情况下保持分散或悬浮于水中,例如以减少水中存在的至少一种病原微生物的量和/或减少或防止水生生物暴露于病原微生物。(Methods of treating water to promote aquatic health are described. The method may comprise introducing the crushed kaolin into water. The crushed kaolin may have a particle size distribution such that at least 30% by weight of the particles pass through a 60 mesh screen. For example, the crushed kaolin can be a kaolin having a d of less than 300 μm 50 In the form of a dry powder of diameter. The pulverized kaolin can be classified intoThe dispersant and/or surfactant, in the case of a dispersant and/or surfactant, remain dispersed or suspended in the water, e.g., to reduce the amount of at least one pathogenic microorganism present in the water and/or to reduce or prevent exposure of the aquatic organism to the pathogenic microorganism.)

1. A method of treating water, the method comprising:

introducing pulverized kaolin into the water, wherein the pulverized kaolin has a particle size distribution such that at least 30% by weight of the particles pass through a 60 mesh sieve, and wherein the pulverized kaolin reduces the amount of at least one pathogenic microorganism present in the water.

2. The method of claim 1, wherein the crushed kaolin remains suspended in the water for at least 2 hours.

3. The method according to claim 1, wherein the crushed kaolin is dispersed in water to a concentration of from about 0.01g/L to about 8.0 g/L.

4. The method of claim 1, where the at least one pathogenic microorganism comprises a bacterium, a protozoan, a virus, a fungus, a parasite, or a combination thereof.

5. The method of claim 1, where the at least one pathogenic microorganism is selected from flavobacterium columnare, edwardsiella ictaluri, edwardsiella tarda, saprolegnia, or combinations thereof.

6. The method according to claim 1, wherein the crushed kaolin comprises less than 0.05% by weight of a dispersant.

7. The method of claim 1, wherein the crushed kaolin is in the form of a dry powder and does not contain a dispersant or surfactant.

8. The method of claim 1, wherein the water comprises at least one aquatic organism selected from the group consisting of fish, shellfish, crustaceans, or combinations thereof, and wherein the method reduces exposure of the at least one aquatic organism to the at least one pathogenic microorganism.

9. The method of claim 1, wherein the at least one aquatic organism comprises a fish selected from the group consisting of catfish, tilapia, carp, salmon, sea bass, eel, mullet, snapper, amber fish, grouper, sea bass, trout, sturgeon, or turbot.

10. The method of claim 1, wherein the water comprises fresh water.

11. The method of claim 1, wherein the crushed kaolin has a d of less than 300 μm₅₀In the form of a dry powder of diameter.

12. A method of treating water, the method comprising:

introducing pulverized kaolin into water, wherein the pulverized kaolin disperses in the water to a concentration of about 0.01g/L to about 8.0g/L within 1 hour;

wherein the water comprises fresh water contained in a mobile tank; and is

Wherein the crushed kaolin reduces the amount of at least one pathogenic microorganism present in the fresh water.

13. The method of claim 12, wherein the volume of water contained in the mobile tank is in the range of about 1,000 gallons to about 5,000 gallons.

14. The method of claim 12, wherein the crushed kaolin is added to the water at least 6 hours before the mobile tank is transported from the first location to the second location.

15. The method of claim 12, wherein the water comprises at least one fish selected from the group consisting of catfish, tilapia, carp, salmon, sea bass, eel, mullet, snapper, amber, grouper, sea bass, trout, sturgeon, turbot, goldfish, koi, croaker, peacock, or combinations thereof.

16. The method of claim 12, where the at least one pathogenic microorganism comprises a bacterium, a protozoan, or a combination thereof.

17. The method of claim 16, where the at least one pathogenic microorganism is selected from flavobacterium columnare, edwardsiella ictaluri, edwardsiella tarda, saprolegnia, or combinations thereof.

18. The method according to claim 12, wherein the crushed kaolin has a particle size distribution such that from about 85% to about 93% by weight of the kaolin particles are retained on a 200 mesh sieve and from about 90% to 100% by weight of the particles are retained on a 325 mesh sieve.

19. The method according to claim 12, wherein the crushed kaolin has a d from about 50 μm to about 350 μm₅₀In the form of a dry powder of diameter.

20. A method of treating water for aquaculture, the method comprising:

introducing pulverized kaolin into water, wherein the pulverized kaolin has a particle size distribution wherein at least 40% by weight of the particles pass through a 60 mesh sieve;

wherein the crushed kaolin is dispersed in water to a concentration of about 0.01g/L to about 8.0 g/L; and is

Wherein the amount of at least one pathogenic microorganism selected from bacteria or protozoa present in the water is reduced by the comminuted kaolin to reduce exposure of fish contained in the water to the at least one pathogenic microorganism.

21. The method of claim 20, wherein the at least one pathogenic microorganism comprises saprolegnia.

22. The method of claim 20, wherein the water comprises at least one juvenile fish selected from the group consisting of catfish, tilapia, carp, salmon, sea bass, eel, mullet, snapper, amber, grouper, river bass, trout, sturgeon, or turbot, and wherein the method increases the survival rate of the at least one juvenile fish.

23. The method of claim 20, wherein the water is contained in a mobile tank, and wherein the crushed kaolin is added to the water at least 12 hours before the mobile tank is transported from the first location to the second location.

Technical Field

The present disclosure relates to compositions and methods for treating water in fish farms and other aquatic environments. More particularly, the present disclosure relates to the use of finely particulate clays (e.g., crushed clays) to promote the health of aquatic organisms.

Background

Certain aquatic pathogens in aquatic environments can threaten the health of aquatic organisms such as fish, shellfish and crustaceans. These pathogens include microorganisms, such as bacteria and protozoa. For example, the column disease is caused by the rod-shaped gram-negative bacterium Flavobacterium columnare (Flavobacterium columnare) As a result, this is an opportunistic pathogen that causes a large number of deaths in freshwater fish species worldwide. Channel catheterus (Channel cathesh) which is economically valuable has been found to be very sensitive to this pathogen. Another example of an aquatic pathogen is Saprolegnia (Saprolegnia), a protozoan that often targets fish and causes skin necrosis.

Despite the importance and impact of the diseases caused by these pathogens, there are few readily available therapeutic or prophylactic measures. Antibiotics have been shown to exhibit some efficacy against cylindrosis, but the use of antibiotics in aquaculture is under increasingly stringent scrutiny and is becoming undesirable. Furthermore, in recent years, the overuse of antibiotics in aquaculture has increased the risk of resistance to aquaculture pathogens (e.g., flavobacterium columnare).

Thus, there is a need for alternative prophylactic and therapeutic approaches, including non-antibiotic based treatments, to diseases caused by pathogens affecting aquaculture.

SUMMARY

The present disclosure includes compositions for promoting the health of aquatic organisms and methods of use thereof. For example, the methods herein can reduce and/or prevent exposure to pathogenic microorganisms/organisms, such as bacteria, protozoa, viruses, fungi, algae, and/or parasites. Additionally or alternatively, the methods herein can promote aquatic health by increasing the turbidity of the water, e.g., to help increase the survival rate of juvenile fish.

For example, according to some aspects of the present disclosure, there is a method of treating water to reduce and/or prevent bacterial infection in aquatic organisms. The method can include introducing kaolin (e.g., a particulate kaolin clay) into the water such that the kaolin reduces the presence of at least one undesirable microorganism/organism (e.g., a pathogenic microorganism, such as a bacterial species and/or protozoa) present in the water, wherein the kaolin is comminuted. The method may include contacting the aquatic organism with the treated water. In some examples, the method may include contacting a microorganism (e.g., a pathogenic microorganism, such as a bacterium and/or a protozoan) with the treated water. For example, microorganisms can be adsorbed onto the surface of kaolin.

The kaolin clay may have a BET surface area of at least 20m²/g or at least 25m²In terms of/g, e.g. at least 30m²/g or at least 40m²(ii) in terms of/g. In some examples, the kaolin clay can have a BET surface area of about 20m²G to about 40m²A,/g, about 25m²G to about 35m²In the range of/g or about 35m²G to about 40m²/g。

The kaolin can comprise a fine particulate kaolin clay. For example, kaolin may be pulverized. According to some aspects of the present disclosure, the kaolin can include a particulate kaolin clay having a particle size distribution such that at least 70% by weight of the kaolin particles have an Equivalent Spherical Diameter (ESD) of less than 2 μm as determined by a sedimentation diagram (Sedigraph). For example, the kaolin can have a particle size distribution such that at least 80%, at least 85%, or at least 90% by weight of the kaolin particles have an ESD of less than 2 μm as determined by a sedimentation diagram.

In another aspect, the kaolin can have a particle size distribution such that at least 25% by weight of the kaolin particles have an ESD of less than 0.25 μm as determined by sedimentation. For example, the kaolin has a particle size distribution such that at least 30%, at least 40%, or at least 50% by weight of the kaolin particles have an ESD of less than 0.25 μm as determined by a sedimentation diagram.

In another aspect, the kaolin can have a combination of shape factor and particle size such that the product of its shape factor times the weight percent of kaolin clay particles having an ESD less than 0.25 μm as determined by a sedimentation plot has a value of at least about 300. For example, the product of the shape factor of the kaolin clay multiplied by the weight percent of kaolin clay particles having an ESD less than 0.25 μm as determined by a sedimentation plot may have a value of at least about 500 or at least about 1000.

In another aspect, the kaolin can have a combination of specific surface area and particle size such that the product of its specific surface area multiplied by the weight percent of kaolin clay particles having an ESD less than 0.25 μm as determined by a sedimentation diagram has a value of at least about 600. For example, the product of the specific surface area of the kaolin clay multiplied by the weight percent of kaolin clay particles having an ESD less than 0.25 μm as determined by a sedimentation plot may have a value of at least about 800 or at least about 1000.

In another aspect, the particulate kaolin clay can be applied to or introduced into the water to be treated to establish a concentration of from about 0.01g/L to about 8.0g/L, such as from about 0.1g/L to about 7.0g/L, from about 0.5g/L to about 6.0g/L, from about 1.0g/L to about 5.0g/L, from about 2.0g/L to about 4.0g/L, or from about 0.5g/L to about 3.0 g/L. In some examples, the kaolin includes less than 0.1% by weight of a dispersant or surfactant, such as less than 0.05% by weight or less than about 0.01% of a dispersant or surfactant. In some examples, the particulate kaolin does not include any dispersants or surfactants.

Water may be treated with the compositions and methods herein to reduce exposure of aquatic organisms to one or more pathogenic microorganisms or pathogenic organisms. For example, the pathogenic microorganisms may include microorganisms selected from the group consisting of Flavobacterium columnare, Edwardsiella ictaluri (S) ((II))Edwardsiella lctaluri) And Edwardsiella tarda (Edwardsiella tarda) At least one undesirable bacterial species. In another example, the pathogenic microorganisms may include protozoa, such as saprolegnia. In another example, the pathogenic microorganisms or pathogenic organisms may include algae.

The aquatic organism may comprise at least one fish, shellfish or crustacean. The fish may in particular comprise, for example, one or more fish selected from catfish, tilapia, carp, barbel and other carps, salmon, sea bass, eel, mullet, sea bream, amber fish, grouper, river bass, trout, sturgeon or turbot. In some examples, the aquatic organism includes at least one crustacean, such as a crustacean selected from the group consisting of shrimp, lobster, crab, or crayfish. Additionally or alternatively, the aquatic organism may include at least one shellfish, such as oysters, scallops, mussels, or clams.

According to some aspects of the present disclosure, a method of treating water may comprise introducing pulverized kaolin into water, wherein the pulverized kaolin may have a particle size distribution such that at least 30% by weight of the particles pass through a 60 mesh sieve, and wherein the pulverized kaolin reduces the amount of at least one pathogenic microorganism present in the water. In some examples, the crushed kaolin can have a particle size distribution such that about 85% to about 93% by weight of the kaolin particles are retained on a 200 mesh sieve and about 90% to 100% by weight of the particles are retained on a 325 mesh sieve. The comminuted kaolin can be in the form of a dry powder, e.g., d₅₀The diameter is less than 300 μm, for example less than 200 μm or less than 100 μm. For example, d of dry powders of comminuted kaolin₅₀The diameter may be in the range of about 50 μm to about 350 μm.

After introducing the crushed kaolin into the water, the crushed kaolin may remain suspended in the water for at least 2 hours, e.g., at least 4 hours or at least 6 hours. In at least one example, the pulverized kaolin is dispersed in water to a concentration of about 0.01g/L to about 8.0g/L, such as about 1g/L to about 5.0 g/L. In at least one example, the dried, pulverized kaolin does not contain dispersants, surfactants, or other chemical additives.

According to some aspects of the present disclosure, water may be contained in a mobile tank. For example, a mobile tank can hold a volume of water of about 1,000 gallons to about 5,000 gallons or about 2,000 gallons to about 4,000 gallons. The water of the portable tank may comprise at least one fish, for example, one or more fish selected from catfish, tilapia, carp, salmon, sea bass, eel, mullet, snapper, amber fish, grouper, perch, trout, sturgeon, turbot, goldfish, koi, croaker (beta), or peacock.

The pulverized kaolin may be added to the portable tank one or more times, such as twice or three times daily, every other day, once a week, or once every two weeks. In at least one example, the crushed kaolin is added to the water at least 3 hours, at least 6 hours, or at least 12 hours prior to transporting the mobile tank from the first site to the second site, such as from the first pond to the second pond, from the pond to a vehicle (e.g., a truck for transporting the tank), or from the vehicle to the pond.

It is to be understood that both the foregoing general description and the following detailed description are exemplary and explanatory only and are not restrictive of the invention.

Brief Description of Drawings

The accompanying drawings, which are incorporated in and constitute a part of this specification, illustrate various exemplary aspects of the disclosure and together with the description, serve to explain the principles of the disclosure.

FIG. 1 is a graph showing the relative observed colony count of Flavobacterium columnare obtained after treatment with each kaolin evaluated in the examples compared with the content of kaolin fine particles having a particle size of less than 0.25 μm as determined by a sedimentation chart.

FIG. 2 is a graph showing the relative observed Flavobacterium columnare colony counts obtained after treatment with each kaolin evaluated in the examples compared to the product of the shape factor of each kaolin multiplied by the% of kaolin fine particles having a particle size of less than 0.25 μm as determined by a sedimentation plot.

FIG. 3 is a graph showing the relative observed fish mortality after exposure to Flavobacterium columnare after treatment with each kaolin evaluated in the examples compared to the BET surface area of each kaolin.

FIG. 4 is a graph showing the relative observed fish mortality after exposure to Flavobacterium columnare after treatment with each kaolin evaluated in the examples compared to the product of the shape factor of each kaolin multiplied by the% of kaolin fines having a particle size of less than 0.25 μm as determined by a sedimentation plot.

FIG. 5 is a graph showing the relative observed fish mortality after exposure to Flavobacterium columnare after treatment with each kaolin evaluated in the examples compared to the product of the specific surface area of each kaolin multiplied by the% of kaolin fines having a particle size of less than 0.25 μm as determined by a sedimentation plot.

Detailed description of the invention

Specific aspects of the disclosure are described in more detail below. To the extent that there is a conflict with a term and/or definition incorporated by reference, the term and definition provided herein shall control.

As used herein, the terms "comprises," "comprising," or any other variation thereof, are intended to cover a non-exclusive inclusion, such that a process, method, composition, article, or apparatus that comprises a list of elements does not include only those elements but may include other elements not expressly listed or inherent to such process, method, composition, article, or apparatus. The term "exemplary" is used in the sense of "example" rather than "ideal".

As used herein, the singular forms "a", "an" and "the" include plural referents unless the context clearly dictates otherwise. The terms "approximately" and "about" mean approximately the same number or value as is referenced. It is understood that the terms "approximately" and "about" as used herein include ± 5% of the specified amount or value.

According to some aspects of the present disclosure, water, such as fine-grained kaolin clay, may be treated with kaolin to promote aquatic organism health, such as the health of fish, crustaceans, and/or shellfish and other aquatic organisms, for example, the methods herein may help reduce and/or prevent exposure of aquatic organisms to pathogenic microorganisms, such as bacteria, protozoa, viruses, fungi, and/or parasites, further, for example, the methods herein may help maintain or increase fish populations, such as by improving the survival rate of juvenile fish, still water (e.g., tanks, streams, bays, etc.) and/or moving water (e.g., streams, bays, etc.) may be treated with the methods herein, water may include fresh water, or in some cases, salt water, water may be contained in fixed containers or enclosures (e.g., ponds or ponds), or in transportable containers or enclosures (e.g., tanks or vats), for example, tanks or vats, such as tank holding tanks or vats, for example, a method herein may use nano 500 gallons (~ liters) to about 5,000 liters (e.g., ~,000 liters) to about 3,000 liters (e.g., 3,800 liters) to about 500 liters, 67000 liters, e.g., 3,800 liters to about 500 liters, and/or more liters, e.g., 3 liters, 500 liters per liter, 9.

In some examples, the method includes introducing the kaolin into the water at a dosage sufficient to reduce the presence of at least one undesirable microorganism (pathogenic microorganism, e.g., bacterial species and/or protozoa) present in the water. The method may further comprise contacting the aquatic organism with the treated water. In some examples, the method can include contacting a microorganism (e.g., a pathogenic microorganism, such as a bacterium and/or a protozoan) with the treated water.

Kaolin comprises mainly the mineral kaolinite (having the general formula Al)₂Si₂O₅(OH)₄Aqueous aluminosilicates) with lesser amounts of various other minerals such as smectite, mica and iron compounds. Exemplary smectite clays include, for example, montmorillonite ((Na, Ca)_0.33(Al,Mg)₂(Si₄O₁₀)(OH)₂•nH₂O), nontronite, beidellite, and saponite. In some aspects of the disclosure, the kaolin can comprise at least 50% by weightKaolinite, for example, from 50 wt% to about 90 wt% kaolinite, or from about 65 wt% to about 75 wt% kaolinite.

The compositions herein may comprise kaolin formulated to facilitate dispersion and/or increase dispersion in the body of water to be treated. For example, the kaolin may comprise a pulverized kaolin, such as a fine particulate kaolin. The pulverized kaolin can provide adequate dispersion without the addition of surfactants or other dispersants, and without any chemical treatment of the kaolin to increase dispersion. Thus, the methods herein can provide compositions with good dispersion characteristics at a lower cost than clays incorporating dispersants. In addition, the crushed kaolin avoids the introduction of dispersants or other chemical additives that may be detrimental to the health of aquatic organisms.

The particle size distribution of the comminuted kaolin can be determined by sieve analysis, for example, using a Rotap shaker. In this procedure, several screens of different mesh sizes are stacked so that the screen holes become smaller in order from top to bottom. The granular sample is placed on the top screen and the instrument then knocks or shakes the stack of screens for a specified period of time (e.g., 10 minutes). This rapping causes the sample to move around on the first screen and material smaller than the mesh falls from the first screen to the next. This continues until the original sample is separated into its various mesh fractions. The results may be expressed as a percentage retained on each sieve or alternatively, as a percentage passed through each sieve. For example, the crushed kaolin may be analyzed in a sieve of 60 mesh (250 μm), 100 mesh (150 μm), 200 mesh (75 μm), and/or 325 mesh (45 μm).

In some aspects of the disclosure, the kaolin can have a particle size distribution such that at least 20% by weight of the particles pass through a 60 mesh screen, e.g., at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, or even at least 55% by weight. For example, about 25 wt% to about 55 wt% or about 40 wt% to about 50 wt% of the particles may pass through a 60 mesh screen.

In some examples, the crushed kaolin can have a particle size distribution such that from about 45% by weight to about 65% by weight, such as from about 50% by weight to about 60% by weight, or about 55% by weight of the particles are retained on a 60 mesh sieve; and/or from about 60 wt% to about 80 wt%, such as from about 65 wt% to about 75 wt%, such as about 70 wt%, of the particles are retained on a 100 mesh screen; and/or from about 80 wt% to about 95 wt% of the particles, such as from about 85 wt% to about 93 wt%, such as about 90 wt% of the particles, are retained on a 200 mesh screen, and/or from about 90 wt% to 100 wt%, such as from about 95 wt% to about 99 wt%, such as about 99 wt% of the particles are retained on a 325 mesh screen.

Another method of characterizing particle size uses SEDIGRAPH instruments (e.g., SEDIGRAPH5100 available from Micromeritics Corporation, USA). In this procedure, the settling rate of the dispersed particles of the sample is determined by a standard dilute aqueous suspension. The size of a given particle may be expressed in terms of the diameter of an equivalent diameter sphere (i.e., "equivalent sphere diameter" or ESD) that settles through the suspension, which may be used to characterize the particulate material. The settlement plot records the weight percent of particles having an ESD less than a particular ESD value, relative to that ESD value.

In some examples, the crushed kaolin can have a particle size distribution such that greater than 75% by weight of the particles, greater than 80% by weight of the particles, greater than 85% by weight of the particles, greater than 90% by weight of the particles, or even greater than 95% by weight of the particles have an ESD of less than 2 μm as determined by sedimentation diagram. In some examples, in terms of fine particle content, the crushed kaolin can have a particle size distribution in which at least 20% by weight of the particles have an ESD less than 0.25 μm, e.g., at least 25%, at least 30%, at least 40%, or at least 50% by weight of the particles have an ESD less than 0.25 μm as determined by sedimentation diagram. In some examples, about 50 wt% to 100 wt%, or about 60 wt% to about 90 wt% of the kaolin particles can have an ESD of less than 0.25 μm as determined by sedimentation.

In some examples, the crushed kaolin can have a particle size distribution such that at least 70% by weight of the kaolin particles have an ESD of less than 2 μm as determined by the sedimentation diagram, e.g., at least 80%, at least 85%, or at least 90% by weight have an ESD of less than 2 μm as determined by the sedimentation diagram. Additionally, in some examples, the crushed kaolin can have a particle size distribution such that at least 25% by weight of the kaolin clay particles have an ESD as determined by the sedimentation diagram of less than 0.25 μm, for example at least 30%, at least 40%, or at least 50% by weight have an ESD as determined by the sedimentation diagram of less than 0.25 μm.

It should be noted that the sedimentation method determines the size of individual particles in a liquid dispersion, while the Rotap method determines coarse aggregates of dry particles, e.g., without the use of surfactants or other dispersants to separate the particles. Thus, the Rotap sieve assay method generally provides a particle size measurement indicative of the dried pulverized material.

The particle size distribution according to the Rotap or sedimentation diagram determination procedure may be expressed in terms of the weight percentage of particles having a size smaller than a particular diameter. E.g. d₅₀At 50% by weight of the particles having a d of less than₅₀The diameter of the size of the value. Similarly, d₃₀Is 30% by weight of the particles have a particle size of smaller size, and d₇₀70% by weight of the particles have a smaller particle size. The term "coarse" generally refers to particles in which less than 30% by weight have a d of less than 0.25 μm₅₀Particle size distribution of diameters, and "fine" generally refers to particles in which greater than 30% by weight of the particles have a d of less than 0.25 μm₅₀Particle size distribution of diameter.

According to some aspects of the present disclosure, the dried, comminuted kaolin clay as determined by Rotap can have a d of less than 300 μm₅₀Particle size, for example less than 250 μm, less than 200 μm, less than 150 μm or less than μm. For example, the dried, comminuted kaolin clay as determined by Rotap can have a d ranging from about 50 μm to about 300 μm, from about 100 μm to about 250 μm, or from about 150 μm to about 200 μm₅₀A diameter, e.g., d, of about 50 μm, about 100 μm, about 150 μm, about 200 μm, about 250 μm, or about 300 μm₅₀Diameter. It should be noted that d measured on the crushed kaolin by the sedimentation plot is expected because the sedimentation plot measures individual dispersed particles rather than the coarser aggregates of the dried crushed kaolin clay₅₀Particle size ratio of dried pulverized kaolin₅₀The particle size is small (e.g., less than about 2 μm).

In some aspects of the present disclosure, the crushed kaolin can have a platy or blocky shape. Relatively high shape coefficient kaolin particles can be considered more "platy" than low shape coefficient kaolin particles, while low shape coefficient kaolin particles can be considered more "blocky". As used herein, "shape factor" is a measure (based on weight average) of the average of the ratio of the average particle diameter to the particle thickness of a population of particles of different sizes and shapes, as determined using the conductivity methods and instruments described in patent publications GB 2,240,398, US 5,128,606, EP 0528078, US 5,576,617, and EP 631665, and using the formulas derived in these publications. For example, in the assay method described in EP 0528078, the conductivity of a fully dispersed aqueous suspension of the particles to be tested is passed through an elongated tube. Conductivity measurements are made between (a) a pair of electrodes spaced apart from each other along the longitudinal axis of the tube and (b) a pair of electrodes spaced apart from each other across the transverse width of the tube, and the form factor of the particulate material being measured is determined by using the difference between the two conductivity measurements. A form factor greater than 30 generally describes a plate-like material, while a form factor less than 30 generally describes a block-like material. "average particle diameter" is defined as the diameter of a circle having the same area as the largest face of the particle.

In some examples, the crushed kaolin can be platy, e.g., having a shape factor greater than 30, greater than 35, greater than 40, greater than 50, greater than 70, or even greater than 100. For example, the crushed kaolin can have a shape factor ranging from 30 to 200, 35 to 150, 40 to 100, or 50 to 70. In other examples, the crushed kaolin can be blocky, e.g., having a shape factor equal to or less than 30, e.g., a shape factor ranging from 1 to 30, 5 to 15, or 3 to 10.

BET surface area refers to a technique for calculating the specific surface area of a physically absorbing molecule according to Brunauer, Emmett, and Teller ("BET") theory. The BET surface area may be determined by any suitable measurement technique. As a non-limiting example, a Gemini III2375 surface area analyzer from Micromeritics Instrument Corporation (Norcross, Georgia, USA) may be used to determine BET surface area using pure nitrogen as the sorbed gas.

According to some aspects of the disclosure, the kaolin clay may have a BET surface area of at least 20m²In g, e.g. at least 25m²A ratio of/g, at least 30m²G orAt least 40m²(ii) in terms of/g. For example, the kaolin clay can have a BET surface area of about 25m²G to about 40m²G, about 30m²G to about 40m²In the range of/g or about 35m²G to about 40mm²/g。

Fine-grained kaolin suitable for use in the compositions and methods herein can be prepared from crude kaolin and/or natural kaolin clay (e.g., obtained directly from a kaolin clay deposit). The natural kaolin clay can include virgin kaolin and/or precipitated kaolin. Virgin kaolin clays are those found in the deposits at the site of their formation. For example, kaolin clays of the predominantly primary type are obtained from mineral deposits in England, France, Germany, Spain and the Czech republic. Sedimentary kaolin clays are those that are washed out of the granite matrix in which they are formed during geological times and are deposited in areas remote from their formation site, typically in basins formed in the surrounding formation. For example, kaolin clays obtained from deposits and brazil in the southeast united states are generally of the sedimentary type.

The comminuted kaolin may facilitate its dispersion in the body of water to be treated, for example, allowing the kaolin to be well distributed throughout the water. As a result, the pulverized kaolin can provide significant benefits in therapeutic efficacy compared to natural kaolin or even crude kaolin provided by limited amounts of grinding. For example, the compositions herein can remain dispersed or suspended in water (such that the water is at least partially turbid) for a period of at least 2 hours, at least 4 hours, at least 6 hours, at least 8 hours, at least 10 hours, or at least 12 hours or more. For example, the pulverized kaolin can remain dispersed or suspended in the water for 2 hours to 24 hours, 4 hours to 18 hours, 6 hours to 12 hours, or 4 hours to 16 hours. In contrast, natural or crude kaolin may not be uniformly distributed in water and/or may settle out of water, which may not be sufficient to reduce or prevent the accumulation of pathogenic microorganisms and/or reduce the occurrence of parental appeals. For example, natural kaolin can settle out of water in less than 1 hour or less than 10 minutes. Similarly, crude kaolin (e.g., wherein greater than 70% by weight of the particles have a d greater than 0.25 μm₅₀Diameter) can remain dispersed or suspended in water for less than 2 hours orLess than 1 hour.

The method of making the comminuted kaolin can be dry, and thus does not involve wet processing. Additionally, for example, the method may not include any chemical treatment steps, e.g., the preparation of the comminuted kaolin may not include treating the natural or raw kaolin with a chemical agent, such as a surface treatment agent, a dispersant, a surfactant, or other chemical agent. According to some aspects of the present disclosure, the natural kaolin or raw kaolin may be subjected to one or more pretreatment steps prior to pulverization. For example, the natural kaolin may be processed in a slicer, shredder, crusher, chipper, deblocker, and/or dryer. In at least one example, the natural kaolin can be pre-treated in one or more of a slicer, shredder, crusher, chipper, grinder, or deblocker, followed by drying. For example, natural kaolin may be dried, for example, oven dried at a temperature of about 100 ℃. In some aspects of the present disclosure, the natural kaolin or crude kaolin may not have any pretreatment prior to pulverization.

The comminution may be carried out with any comminution apparatus suitable for providing the finely divided kaolin disclosed herein. In some examples, the pulverization can be carried out with a mill such as a hammer mill, a ball mill, a roller mill, an impact mill, or an air classification mill. According to some aspects of the present disclosure, the crushed kaolin can have a moisture content of less than 3.0% by weight, such as less than 2.0% by weight, less than 1.5% by weight, or less than 1.0% by weight. For example, the moisture content of the comminuted kaolin can be in a range from about 0.1 wt% to about 1.5 wt%, from about 0.5 wt% to about 1.2 wt%, e.g., a moisture content of about 0.5 wt%, about 0.6 wt%, about 0.7 wt%, about 0.8 wt%, about 0.9 wt%, about 1.0 wt%, about 1.1 wt%, about 1.2 wt%, about 1.3 wt%, about 1.4 wt%, or about 1.5 wt%.

The fine particle size of the comminuted kaolin can allow the kaolin to disperse and remain suspended as water is introduced. Thus, for example, the composition may comprise less than 0.1%, less than 0.05%, or less than 0.01% by weight of the dispersant. In some examples, the composition may not include any dispersant. For example, according to some aspects of the present disclosure, a composition added to water for treatment may consist of or consist essentially of comminuted kaolin.

Still water (e.g., ponds, lakes, wetlands, tanks, etc.) and/or running water (e.g., streams, bays, rivers, etc.) may be treated with the composition of comminuted kaolin. For example, the water may comprise fresh water. Additionally, water contained in natural water bodies and/or man-made structures (e.g., artificial lakes) may be treated with the compositions and methods herein. The water may comprise one or more species of fish, crustaceans and/or shellfish and other aquatic organisms.

The composition of comminuted kaolin can be applied to water to establish a concentration of kaolin in a range of about 0.01g/L to about 8.0g/L, such as about 0.1g/L to about 7.0g/L, 0.5g/L to about 6.0g/L, about 1.0g/L to about 5.0g/L, about 2.0g/L to about 4.0g/L, or about 0.5g/L to about 3.0g/L, about 0.1g/L to about 2.0g/L, or about 1.0g/L to about 5.0 g/L. For example, the pulverized kaolin can be added to water (e.g., water contained in a vat or tank, such as a mobile vat or tank) as a dry powder that is dispersible in water to achieve a concentration of about 0.1g/L to about 8/0g/L over a period of about 5 minutes to about 3 hours, such as a period of about 10 minutes to about 1 hour, such as about 20 minutes, about 30 minutes, about 40 minutes, or about 1 hour.

The pulverized kaolin can be introduced into the water one or more times to increase or maintain the desired concentration of kaolin suspended or otherwise placed in the water. For example, the crushed kaolin can be introduced into the water once a day, twice a day, three times a day, once a week, twice a week, once every two weeks (two weeks), once a month, or with other timing schemes, e.g., based on the volume of the water, the quality of the water (e.g., concentration and/or chemical composition of suspended solids), the type of aquatic organisms present in the water, and/or the amount of aquatic organisms present in the water.

According to some aspects of the present disclosure, the crushed kaolin may be added to the water prior to anticipating the transport of fish or other aquatic organisms contained in the water from the first habitat to the second habitat. For example, the pulverized kaolin may be added to the new habitat to which the fish are transferred before, during, or immediately after the fish are transferred to the new habitat. Similarly, the pulverized kaolin can be added to the water of the original habitat before the fish are expected to be transferred to the new habitat (e.g., one, two, or three or more days before). In at least one example, the pulverized kaolin can be added to a mobile tank or vat into which the fish are introduced during transfer of the fish between bodies of fixed water (e.g., a first fixed tank to a second fixed tank). Additionally, for example, the crushed kaolin can be added to the body of water one or more times (including, e.g., periodically) in addition to and/or independent of the intended transfer of aquatic life contained in the water.

Without intending to be limited by theory, it is believed that the kaolin particles may provide a surface to which pathogenic microorganisms and/or pathogenic organisms may be adsorbed. As kaolin settles over time onto the bed at the bottom of the water, kaolin can effectively remove pathogens from the water, thereby reducing contact with aquatic organisms such as fish, crustaceans, and/or shellfish living in the water.

Water may be treated with the compositions and methods herein to reduce and/or prevent exposure of aquatic organisms to pathogenic microorganisms/organisms selected from the group consisting of: bacteria (including, for example, gram negative bacteria), protozoa, viruses, fungi, algae, and/or parasites. Exemplary pathogenic microorganisms include, but are not limited to, Flavobacterium columnare (F.) (Flavobacterium columnare) Edwardsiella ictaluri (C), Edwardsiella ictaluri (C)Edwardsiella lctaluri) Edwardsiella tarda (Edwardsiella tarda) Saprolegnia parasitica (B)Saprolegnia) Aeromonas salmonicida (Aeromonas salmonicida) Aeromonas hydrophila (b) ((b))Aeromonas hydrophila)、Aeromonas formicansAeromonas liquefaciens (Aeromonas liquefaciens) Aeromonas hydrophila (b) ((b))Aeromonas hydrophila) Yersinia ruckeri: (Yersinia ruckeri) Salmonella bacteria (A), (B), (C)Renibacterium salmoninarum) Flavobacterium psychrophilum (a)Flavobacterium psychrophilum) Cytophaga bacterium (a)Cytophaga) Flavobacterium gillonum (f.) KuntzeFlavobacterium branchiophila) And Vibrio harveyi: (Vibrio harveyi). In some examples, the water comprises at least one gram-negative bacterium. In at least one example, the pathogenic microorganisms present in the water include flavobacterium columnare. In at least one example, a pathogenic microorganism (or etiology)Organisms) may include algae.

The water to be treated may comprise one or more types of aquatic organisms, such as one or more fish, shellfish and/or crustaceans. According to some aspects of the disclosure, the aquatic organism comprises at least one fish, such as a fish selected from catfish, tilapia, carp, salmon, sea bass, eel, mullet, snapper, amber fish, grouper, river bass, trout, sturgeon, or turbot, among others. In some examples, the fish may include ornamental fish, such as goldfish, koi, croaker, and/or malachite. For example, the fish may be contained in a koi pond, aquarium or aquatic waterscape. Additionally or alternatively, the aquatic organism may comprise at least one crustacean, such as a crustacean selected from the group consisting of shrimp, lobster, crab, or crayfish. Additionally, or alternatively, the aquatic organism may include at least one shellfish, such as oysters, scallops, mussels, or clams.

According to some aspects of the present disclosure, the composition may be introduced into water in an amount sufficient to increase the turbidity of the water for a period of time of from 2 hours to 24 hours or more. Increasing the turbidity of water by introducing the compositions of comminuted kaolin disclosed herein can increase the survival rate of juvenile fish (e.g., fingerlings and fry). Without intending to be limited by theory, it is believed that increasing the turbidity of water may limit the ability of the fish to localize and feed on juvenile fish, including, for example, parental diets. For example, according to some aspects of the present disclosure, kaolin can increase the survival rate of juvenile fish.

Other than in the examples, or where otherwise indicated, all numbers expressing quantities of ingredients, reaction conditions, and so forth used in the specification and claims are to be understood as being modified in all instances by the term "about". Accordingly, unless indicated to the contrary, the numerical parameters set forth in the specification and attached claims are approximations that may vary depending upon the desired properties sought to be obtained by the present disclosure. At the very least, and not as an attempt to limit the application of the doctrine of equivalents to the scope of the claims, each numerical parameter should at least be construed in light of the number of reported significant digits and by applying ordinary rounding techniques.

Notwithstanding that the numerical ranges and parameters setting forth the broad scope of the invention are approximations, the numerical values set forth in the specific examples are reported as precisely as possible unless otherwise indicated. Any numerical value, however, inherently contains certain errors necessarily resulting from the standard deviation found in their respective testing measurements.

Examples of certain embodiments of the present disclosure are given below by way of non-limiting illustration.

Examples

The following examples evaluate the efficacy of six different kaolin clays on flavobacterium pillared when used to treat water. The particle size and shape coefficient characteristics of the six kaolins tested are summarized in table 1 below. The particle size information provides a weight percentage of particles having a diameter less than a specified diameter (ESD).

TABLE 1

Sample number	< 2 µm	< 0.5 µm	< 0.25 µm	Form factor
					Kaolin 1	60.5%	27.0%	12.9%	4.6
Kaolin 2	90.3%	76.5%	50.1%	8.4
					Kaolin 3	55.5%	30.5%	14.7%	23.2
Kaolin 4	85.8%	66.7%	45.5%	27.4
					Kaolin 5	62.4%	32.1%	13.8%	8.2
Kaolin 6	48.6%	16.9%	5.5%	7.4

Twenty channel catfish fries (each weighing about 5g on average) were placed in an 18L tank containing 10L of filtered water. Water was provided at a rate of 29.1mL/min by an ultra low flow water delivery system. No fish was fed on the first day after challenge, but pelleted catfish feed (35% protein, 2.5% fat; Delta Western) was provided on the second day and for the remainder of the study.

Fish were experimentally challenged with the virulent flavobacterium columnare isolate LSU-066. Isolates were retrieved from glycerol stocks stored at 80 ℃ and streaked onto Ordal media (Anacker & Ordal 1959). After 48 hours, isolates were removed from agar using sterile cotton swabs and inoculated into 5mL Flavobacterium columnare growth medium (FCGM; Farmer 2004). This suspension was incubated at 28 ℃ for 24 hours and used to inoculate 1L of FCGM. Incubating the inoculated 1L broth in an orbital shaker incubator set at 200rpm for 24 hours at 28 ℃; when the bacteria grew to an absorbance of 0.75 at 550nm, the flask was removed and placed on a stir plate at room temperature. The fish were challenged by adding 5mL of stock bacterial solution to each 10L tank and the calculated exposure dose was 6.2 x 106 CFU/mL. The fish were observed twice daily to assess mortality.

In an exemplary kaolin treatment, 1g/L kaolin was slowly added to the water near the air stones to facilitate mixing in the tank. In the kaolin treated tank, kaolin was added to the water 5 minutes before challenge with flavobacterium columnare to allow sufficient mixing time and initiate ultra low flow. The concentration of kaolin is selected based on previous reports demonstrating that rainbow trout can tolerate the dose well. The duration of the challenge experiment was 7 days.

In another in vitro completed test, a kaolin sample was added to water containing about 9000mio units/liter of flavobacterium columnare. After allowing sufficient mixing time, the kaolin was removed by centrifugation and the supernatant was counted for residual bacteria. The lower the bacterial count in the supernatant, the more effective the removal of bacteria from the kaolin sample. As shown in fig. 1, the relative observed flavobacterium columnare colony counts obtained after treatment with each kaolin evaluated varied greatly according to the particle size of the kaolin. In particular, it has been surprisingly observed that kaolin with a high content of very fine particles having a particle size of less than 0.25 μm shows a higher efficacy against Flavobacterium columnare.

As shown in fig. 2, there is an even greater correlation in observing flavobacterium columnare colony counts when the shape coefficient is considered in addition to the fine particle content by multiplying the shape coefficient of kaolin by its fine particle content (<0.25 μm content). The inhibition of efficacy against Flavobacterium columnare surprisingly high when water is treated with a very fine platy kaolin clay, as shown by the extremely low colony count after treatment with Kaolin 4.

As shown in fig. 3, it was also found that the efficacy against flavobacterium columnare in colony counting experiments was correlated with a reduction in fish mortality when fine kaolin clay was used. FIG. 3 is a graph of the relative observed fish mortality after exposure to Flavobacterium columnare after treatment with each kaolin, compared to the BET surface area of each kaolin. The BET surface area is inversely related to the kaolin particle size, so that a higher BET surface area kaolin corresponds to the same fine particle sample that showed efficacy in colony counting experiments.

As shown in fig. 4, it was also found that the efficacy against flavobacterium columnare in fish mortality experiments was correlated with a decrease in fish mortality when using fine platy clays. Fig. 4 is a graph illustrating the relative observed fish mortality after exposure to flavobacterium columnare after treatment with each kaolin, compared to the product of the shape factor and% fine particles for each kaolin. Fig. 5 is a graph illustrating the relative observed fish mortality after exposure to flavobacterium columnare after treatment with each kaolin, compared to the product of the specific surface area and% fine particles of each kaolin.

Other aspects and embodiments of the disclosure will be apparent to those skilled in the art from consideration of the specification and practice of the embodiments disclosed herein.

It is intended that the specification and examples be considered as exemplary only, with a true scope and spirit of the disclosure being indicated by the following claims.