Description:DESCRIPTION:
Field of the invention:
The present disclosure generally relates to the technical field of an aquaculture, in specific, relates to a composition that exhibits anti-ulcer activity in aquatic species using a Kappaphycus alvarezii powder and a Vetiver powder.
Background of the invention:
The field of ulcer treatment has explored a range of natural and synthetic compounds for their potential to mitigate gastric ulcers and associated symptoms. A significant amount of research has focused on testing various bioactive compounds and formulations to determine their effectiveness in promoting ulcer healing and preventing the recurrence of these lesions. Notably, studies have reported the use of the pyloric ligation model in mice to assess antiulcer activity. This model, widely employed in scientific studies, is instrumental in inducing gastric ulcers for evaluating potential therapeutic agents. Similarly, the ex vivo chick stomach model has been utilized to study the antiulcer properties of bioactive compounds, that include extracts derived from marine sources such as Sargassum ilicifolium.

Research into the bioactive potential of marine algae has shown promising results, particularly in the context of antiulcer activity. Fucoidans, a type of polysaccharide derived from brown algae, have been recognized for their ability to promote ulcer healing and offer protective effects against gastric lesions. Moreover, the exploration of marine-derived bioactive compounds, as seen in the chemical characterization of algae, has opened up avenues for discovering new natural therapies for gastrointestinal disorders. Despite these advances, significant gaps remain in the understanding and utilization of specific marine algae, such as Kappaphycus alvarezii, for antiulcer treatments.

Environmental factors, particularly water pollution, have also been identified as contributors to ulcer formation in various aquatic species, underscoring the potential link between environmental stressors and gastric lesions. While this phenomenon has been documented in fish species, it raises broader questions about the impact of environmental toxins on ulcer development in humans and animals.

Despite the promising results from these studies, there are notable drawbacks associated with the existing methods and models used in antiulcer research. Many of these approaches, such as the chick stomach model, are associated with adverse effects and limited efficacy. Moreover, the focus has largely been on chemical-based methods or conventional treatments, which often fail to incorporate a holistic view that synthesizes ecological knowledge with modern scientific advancements.

The potential of natural compounds such as Kappaphycus alvarezii and Vetiver powder for the antiulcer treatment has been underexplored, leaving a significant gap in the scientific literature. Existing research has yet to fully harness the synergistic potential of these natural compounds, and as a result, current treatments do not maximize their therapeutic benefits. There is a clear and urgent need for a new approach that integrates interdisciplinary scientific advancements to address the shortcomings of existing treatments and develop more effective, natural alternatives.

Addressing these gaps calls for a novel method that combines the therapeutic potentials of Kappaphycus alvarezii and Vetiver powder. By leveraging these natural resources, it is possible to create an innovative formulation that not only offers antiulcer benefits but also promotes sustainability by utilizing ecologically sourced ingredients. This approach represents a transformative shift in the field, promising enhanced efficacy and a broader application of natural compounds in the ulcer treatment.

By addressing all the above-mentioned problems, there is a need for a composition that integrates Kappaphycus alvarezii and Vetiver powder, produced through a double-drying protocol and high-precision multi-stage grinding, to achieve an ultra-fine powder with exceptional uniformity and bioactivity. There is also a need for a cost-effective and sustainable method for preparing the Kappaphycus alvarezii and Vetiver powder formulation, requiring minimal resources and accessible to individuals with limited technical expertise. There is also a need for a method that demonstrates the exceptional efficacy of the Kappaphycus alvarezii and Vetiver formulation in completely healing ethanol-induced gastric ulcers in fish, outperforming current treatments used in aquaculture.

There is also a need for an eco-friendly, chemical-free method for producing the formulation, enhancing ecological resilience and sustainability in aquaculture practices. There is also a need for a Kappaphycus alvarezii and Vetiver formulation underscores its broad-spectrum potential as a natural remedy for ulcers in aquaculture, effectively addressing common ailments worsened by environmental stressors. Further, there is also a need for a formulation that enhances palatability for fish, ensuring greater acceptance and easier administration compared to other treatments, such as antibiotics, which carry the risk of promoting antibiotic resistance.
Objectives of the invention:
The primary objective of the present invention is to provide a composition that exhibits anti-ulcer activity in aquatic species using a Kappaphycus alvarezii powder and a Vetiver powder.

Another objective of the present invention is to provide a composition that integrates Kappaphycus alvarezii and Vetiver powder, produced through a double-drying protocol and high-precision multi-stage grinding, to achieve an ultra-fine powder with exceptional uniformity and bioactivity.

The other objective of the present invention is to provide a cost-effective and sustainable method for preparing the Kappaphycus alvarezii and Vetiver powder formulation, requiring minimal resources and accessible to individuals with limited technical expertise.

The other objective of the present invention is to provide a method that demonstrates the exceptional efficacy of the Kappaphycus alvarezii and Vetiver formulation in completely healing ethanol-induced gastric ulcers in fish, outperforming current treatments used in aquaculture.

The other objective of the present invention is to provide an eco-friendly, chemical-free method for producing the formulation, enhancing ecological resilience and sustainability in aquaculture practices.

Yet another objective of the present invention is to provide to a formulation that enhances palatability for fish, ensuring greater acceptance and easier administration compared to other treatments, such as antibiotics, which carry the risk of promoting antibiotic resistance.

Further objective of the present invention is to provide a Kappaphycus alvarezii and Vetiver formulation underscores its broad-spectrum potential as a natural remedy for ulcers in aquaculture, effectively addressing common ailments worsened by environmental stressors.
Summary of the invention:
The present disclosure proposes a composition and method for treating ulcers in aquatic species using Kappaphycus alvarezii and Vetiver powder. The following presents a simplified summary in order to provide a basic understanding of some aspects of the claimed subject matter. This summary is not an extensive overview. It is not intended to identify key/critical elements or to delineate the scope of the claimed subject matter. Its sole purpose is to present some concepts in a simplified form as a prelude to the more detailed description that is presented later.

In order to overcome the above deficiencies of the prior art, the present disclosure is to solve the technical problem to provide a composition that exhibits anti-ulcer activity in aquatic species using a Kappaphycus alvarezii powder and a Vetiver powder.

According to one aspect, the invention provides a primary composition exhibits anti-ulcer activity. In one embodiment herein, the primary composition comprises 48 to 52 weight percentage of a Kappaphycus alvarezii powder as an active ingredient. The Kappaphycus alvarezii powder is derived from freshly harvested Kappaphycus alvarezii material, which is washed, sun-dried, and ground upon undergoing the dual-phase drying protocol. The Kappaphycus alvarezii powder is prepared through the dual-phase drying protocol followed by the multi-stage grinding operation.

In one embodiment herein, the primary composition further comprises 48 to 52 weight percentage of a Vetiver powder as a natural ingredient. The Vetiver powder is derived from Vetiveria zizanoides root’s material, which are washed, dried, and ground upon undergoing the dual-phase drying protocol. The Vetiver powder is prepared through the dual-phase drying protocol followed by the multi-stage grinding operation.

In one embodiment herein, the dual-phase drying protocol includes an initial natural solar drying phase for at least 24 hr followed by a controlled thermal drying phase at a temperature of at least 70°C for a time period of at least 24 hr, thereby ensuring complete dehydration while preserving the structural integrity and active constituents of the Kappaphycus alvarezii powder and Vetiver powder. The Kappaphycus alvarezii powder and Vetiver powder are obtained through a mechanical grinding and triple-stage micro-sieving process using micro sieves, ensuring uniform particle size distribution and minimizing thermal degradation during the refinement process.

In one embodiment, the Kappaphycus alvarezii powder and the Vetiver powder are blended to obtain a homogeneous powder blend with optimal particle size distribution. The Kappaphycus alvarezii powder and the Vetiver powder are blended at a ratio of at least 1:1, selected based on empirical data supporting synergistic effects that enhance the performance of the primary composition.

In one embodiment, the homogeneous powder blend is dispersed in distilled water, at varying concentrations to prepare the primary composition for precise dosing and application requirements. The primary composition exhibits anti-ulcer activity by promoting the healing of ethanol-induced gastric ulcers in aquatic species. The primary composition is free of chemical additives, providing a natural, eco-friendly solution.

In another embodiment herein, the homogeneous powder blend is dispersing in the distilled water with carboxymethyl cellulose (CMC) at varying concentrations of 25%, 50%, and 75% to prepare as a secondary composition for precise dosing and application requirements. The homogeneous powder blend is dispersing in the distilled water with the carboxymethyl cellulose (CMC), which ensures stable dispersion of the active components, facilitating consistent and uniform interaction with aquatic subjects, such as the fish species (Labeo rohita).

According to another aspect, the invention provides a method for preparing the primary composition. At one step, the Kappaphycus alvarezii powder and Vetiver powder are prepared through a dual-phase drying protocol followed by multi-stage grinding operation by using the Kappaphycus alvarezii material, Vetiveria zizanoides root’s material and micro sieves. At one step, the Kappaphycus alvarezii powder and the Vetiver powder are blended to obtain a homogeneous powder blend with optimal particle size distribution. At one step, the homogeneous powder blend is dispersed in the distilled water at varying concentrations to prepare the primary composition for precise dosing and application requirements.

Further, objects and advantages of the present invention will be apparent from a study of the following portion of the specification, the claims, and the attached drawings.
Detailed description of drawings:
The accompanying drawings, which are incorporated in and constitute a part of the specification, illustrate an embodiment of the invention, and, together with the description, explain the principles of the invention.

FIG. 1A illustrates a block diagram for preparing a Kappaphycus alvarezii powder, in accordance to an example embodiment of the invention.

FIG. 1B illustrates a block diagram for preparing a Vetiver powder, in accordance to an example embodiment of the invention.

FIG. 2A illustrates a schematic representation of a primary composition for exhibiting anti-ulcer activity in aquatic species, in accordance to an example embodiment of the invention.

FIG. 2B illustrates a schematic representation of a secondary composition for exhibiting anti-ulcer activity in aquatic species, in accordance to an example embodiment of the invention.

FIG. 3 illustrates a flowchart of a method for preparing and evaluating the primary composition, in accordance to an example embodiment of the invention.

FIG. 4 illustrates a pictorial representation of aquatic species with ethanol-induced gastric ulcers, in accordance to an example embodiment of the invention.

FIG. 5 illustrates the pictorial representation of the aquatic species upon healing the ethanol-induced gastric ulcers, in accordance to an example embodiment of the invention.

FIG. 6 illustrates a bar chart representation of an efficiency of the primary composition, in accordance to an example embodiment of the invention.

FIG. 7 illustrates a flowchart of a method for preparing the primary composition, in accordance to an example embodiment of the invention.
Detailed invention disclosure:
Various embodiments of the present invention will be described in reference to the accompanying drawings. Wherever possible, same or similar reference numerals are used in the drawings and the description to refer to the same or like parts or steps.

The present disclosure has been made with a view towards solving the problem with the prior art described above, and it is an object of the present invention to provide a primary composition that exhibits anti-ulcer activity in aquatic species using a Kappaphycus alvarezii powder and a Vetiver powder.

According to one example embodiment of the invention, FIG. 1A refers to a block diagram for preparing a Kappaphycus alvarezii powder 102 used in a primary composition 100. In one embodiment herein, the process begins with the systematic collection of freshly harvested Kappaphycus alvarezii material 10 to ensure optimal quality and consistency in their physicochemical properties. The Kappaphycus alvarezii material 10 is carefully selected based on strict criteria to maintain uniformity in their structure, chemical composition, and active constituents.

Once harvested, the Kappaphycus alvarezii material 10 undergo a dual-phase drying protocol to ensure complete dehydration while preserving their integrity. The first phase involves 24 hr of natural solar drying, allowing for initial moisture removal under ambient conditions. This is followed by the second phase, a controlled thermal drying process, where the Kappaphycus alvarezii material 10 is exposed to a temperature of at least 70°C for an additional time period of 24 hr. This controlled environment ensures complete dehydration without compromising the structural integrity or the preservation of the active components, which is crucial for the effectiveness of the primary composition 100.

Following the drying process, the Kappaphycus alvarezii material 10 is prepared for powder production and refinement. The dried Kappaphycus alvarezii are ground using a mechanical grinder equipped with precision-engineered blades designed to minimize heat generation during the grinding process, thus preventing any thermal degradation of sensitive bioactive compounds. The resulting coarse powder is then subjected to a triple-stage micro-sieving process, where sieves 14 calibrated to different mesh sizes ensure uniformity in particle size distribution. This process results in the Kappaphycus alvarezii powder 102 that is ideal for further formulation.

According to one example embodiment of the invention, FIG. 1B refers to a block diagram for preparing a Vetiver powder 104. In one embodiment herein, the process begins with the systematic collection of Vetiveria zizanoides root’s material 12 to ensure optimal quality and consistency in their physicochemical properties. The Vetiveria zizanoides root’s material 12 is carefully selected based on strict criteria to maintain uniformity in their structure, chemical composition, and active constituents.

Once harvested, the Vetiveria zizanoides root’s material 12 undergo the dual-phase drying protocol to ensure complete dehydration while preserving their integrity. The first phase involves 24 hr of natural solar drying, allowing for initial moisture removal under ambient conditions. This is followed by the second phase, a controlled thermal drying process, where the Vetiveria zizanoides root’s material 12 is exposed to a temperature of at least 70°C for an additional 24 hours. This controlled environment ensures complete dehydration without compromising the structural integrity or the preservation of the active components, which is crucial for the effectiveness of the primary composition 100.

Following the drying process, the Vetiveria zizanoides root’s material 12 is prepared for powder production and refinement. The dried Vetiveria zizanoides roots are ground using a mechanical grinder equipped with precision-engineered blades designed to minimize heat generation during the grinding process, thus preventing any thermal degradation of sensitive bioactive compounds. The resulting coarse powder is then subjected to a triple-stage micro-sieving process, where sieves 14 calibrated to different mesh sizes ensure uniformity in particle size distribution. This process results in the Vetiver powder 104 that is ideal for further formulation.

According to one example embodiment of the invention, FIG. 2A refers to a schematic representation of the primary composition 100 for exhibiting anti-ulcer activity in aquatic species. In one embodiment herein, the primary composition 100 comprises 48 to 52 weight percentage of the Kappaphycus alvarezii powder 102 as an active ingredient. The Kappaphycus alvarezii powder 102 is derived from freshly harvested Kappaphycus alvarezii material 10, which is washed, sun-dried, and ground upon undergoing the dual-phase drying protocol. The Kappaphycus alvarezii powder 102 is prepared through the dual-phase drying protocol followed by the multi-stage grinding operation.

In one embodiment herein, the primary composition 100 comprises 48 to 52 weight percentage of the Vetiver powder 104 as a natural ingredient. The Vetiver powder 104 is derived from Vetiveria zizanoides root’s material 12, which are washed, dried, and ground upon undergoing the dual-phase drying protocol. The Vetiver powder 104 is prepared through the dual-phase drying protocol followed by the multi-stage grinding operation.

In one embodiment herein, the dual-phase drying protocol includes an initial 24-hour natural solar drying phase followed by a controlled thermal drying phase at a temperature of at least 70°C for a time period of 24 hr, thereby ensuring complete dehydration while preserving the structural integrity and active constituents of the Kappaphycus alvarezii powder 102 and Vetiver powder 104. The Kappaphycus alvarezii powder 102 and the Vetiver powder 104 are obtained through a mechanical grinding and triple-stage micro-sieving process using micro sieves 14, ensuring uniform particle size distribution and minimizing thermal degradation during the refinement process.

In one embodiment herein, the Kappaphycus alvarezii powder 102 and the Vetiver powder 104 are blended to obtain a homogeneous powder blend with optimal particle size distribution. The Kappaphycus alvarezii powder 102 and the Vetiver powder 104 are blended at a ratio of at least 1:1, selected based on empirical data supporting synergistic effects that enhance the performance of the primary composition 100.

In one embodiment herein, the homogeneous powder blend is dispersed in distilled water at varying concentrations, to prepare the primary composition 100 for precise dosing and application requirements. The homogeneous powder blend is dispersing in the distilled water at varying concentrations of 25%, 50%, and 75% to prepare the primary composition 100 for precise dosing and application requirements. The primary composition 100 exhibits anti-ulcer activity by promoting the healing of ethanol-induced gastric ulcers in aquatic species.

According to another example embodiment of the invention, FIG. 2B refers to the schematic representation of a secondary composition 200 for exhibiting the anti-ulcer activity in the aquatic species. In one embodiment herein, the secondary composition 200 comprises 48 to 52 weight percentage of the Kappaphycus alvarezii powder 102 as the active ingredient. The Kappaphycus alvarezii powder 102 is derived from freshly harvested Kappaphycus alvarezii material 10, which is washed, sun-dried, and ground upon undergoing the dual-phase drying protocol. The Kappaphycus alvarezii powder 102 is prepared through the dual-phase drying protocol followed by the multi-stage grinding operation.

In one embodiment herein, the secondary composition 200 further comprises 48 to 52 weight percentage of the Vetiver powder 104 as the natural ingredient. The Vetiver powder 104 is derived from Vetiveria zizanoides root’s material 12, which are washed, dried, and ground upon undergoing the dual-phase drying protocol. The Vetiver powder 104 is prepared through the dual-phase drying protocol followed by the multi-stage grinding operation.

In one embodiment herein, the dual-phase drying protocol includes an initial 24-hour natural solar drying phase followed by a controlled thermal drying phase at a temperature of at least 70°C for a time period of 24 hr, thereby ensuring complete dehydration while preserving the structural integrity and active constituents of the Kappaphycus alvarezii powder 102 and Vetiver powder 104. The Kappaphycus alvarezii powder 102 and the Vetiver powder 104 are obtained through the mechanical grinding and triple-stage micro-sieving process using micro sieves 14, ensuring uniform particle size distribution and minimizing thermal degradation during the refinement process.

In one embodiment herein, the Kappaphycus alvarezii powder 102 and the Vetiver powder 104 are blended to obtain the homogeneous powder blend with optimal particle size distribution. The Kappaphycus alvarezii powder 102 and the Vetiver powder 104 are blended at a ratio of at least 1:1, selected based on empirical data supporting synergistic effects that enhance the performance of the secondary composition 200.

In one example embodiment herein, the homogeneous powder blend is dispersed in distilled water with carboxymethyl cellulose (CMC) as a vehicle, at varying concentrations of 25%, 50%, and 75% to prepare the secondary composition 200 for precise dosing and application requirements. The homogeneous powder blend is dispersing in the distilled water with the carboxymethyl cellulose (CMC), which ensures stable dispersion of the active components, facilitating consistent and uniform interaction with aquatic subjects, such as the fish species (Labeo rohita).

According to another example embodiment of the invention, FIG. 3 refers to a flowchart 300 of a method for preparing and evaluating the primary composition 100. At step 302, the systematic collection of freshly harvested Kappaphycus alvarezii material 10 and the Vetiveria zizanoides root’s material 12 to ensure optimal quality and consistency in their physicochemical properties. The Kappaphycus alvarezii material 10 and the Vetiveria zizanoides root’s material 12 are carefully selected based on strict criteria to maintain uniformity in their structure, chemical composition, and active constituents.

At step 304, the Kappaphycus alvarezii material 10 and the Vetiveria zizanoides root’s material 12 undergo the dual-phase drying protocol. The first phase involves 24 hours of natural solar drying, followed by the second phase, where the materials (10, 12) are subjected to controlled thermal drying at a temperature of at least 70°C for an additional 24 hr. This process ensures complete dehydration, preserving the structural integrity and active constituents.

At step 306, The dried Kappaphycus alvarezii material and Vetiveria zizanoides roots are ground using a mechanical grinder equipped with precision-engineered blades designed to minimize heat generation during the grinding process, thus preventing any thermal degradation of sensitive bioactive compounds. The resulting coarse powder is then subjected to a triple-stage micro-sieving process, where sieves 14 calibrated to different mesh sizes ensure uniformity in particle size distribution. This process results in the Kappaphycus alvarezii powder 102 and the Vetiver powder 104 that are ideal for further formulation.

At step 308, the Kappaphycus alvarezii powder 102 and the Vetiver powder 104 are meticulously blended in a 1:1 ratio under controlled environmental conditions to obtain the homogeneous powder blend with optimal particle size distribution. This ratio is selected based on empirical data indicating optimal synergistic interactions between the components. At step 310, the multi stage sieving is performed for the homogeneous powder blend using the micro sieves 14. At step 312, The homogenous powder is dispersed in the distilled water to prepare the primary composition 100 of varying concentrations of 25%, 50%, and 75%. The concentrations are precisely calculated to meet the specific requirements of the intended application. The use of Carboxymethyl Cellulose (CMC) as a vehicle ensures consistent suspension of the powder, facilitating uniform distribution during the application. The powered formulation of the primary composition 100 is obtained upon performing the multi stage sieving.

At step 314, the efficiency testing process of the primary composition 100 is initiated. At step 316, the process of gradually adjusting the fishes to a new environment or experimental conditions over a period of 20 days. This ensures that the fish can adapt to factors such as temperature, water quality, and other variables, minimizing stress and ensuring accurate experimental results. At step 318, the ulcer inducing process is initiated. In one embodiment herein, three distinct groups, each of the group consisting of 33 Labeo rohita fishes, classified into control, ranitidine, and test groups. The test group is a powder group, further divided based on the concentration (for example, 25%, 50%, 75%) of the Kappaphycus alvarezii and Vetiver compound administered.

In one embodiment herein, the control group comprises 33 fishes (Labeo rohita), maintained under standardized conditions, receiving no exposure to the formulation. The ranitidine group of 33 fishes is designated, receiving a carefully measured standard compound to serve as a comparative benchmark against the powder formulation. The test group of 33 fishes is constructed, receiving the Kappaphycus alvarezii powder 102 and Vetiver powder 104 in varying concentrations (for example, 25%, 50%, 75%). Each of the concentration is prepared using the afore mentioned protocol to ensure precise and reproducible outcomes.

In one embodiment herein, ethanol solutions are prepared in strict volumetric ratios of 1:3, 1:1, 3:1 to make specific controlled environments in test subjects. This step is critical for standardizing the experimental conditions and enabling the assessment of the powder's efficiency. The ethanol solutions are administered across test and control groups, ensuring uniform exposure. Observations and documentation are conducted systematically, focusing on the precision and consistency of the outcomes.

At step 320, 3 fishes are sacrifice from the each of the group of 33 fishes for documentation of the ulcer and determining effective concentrations. At step 322, the 3 fishes are sacrifice after 7days to observe efficiency. At step 324, the results evaluation process of the primary composition 100 is initiated. In one embodiment herein, the fishes are sacrificed, and gastric juice is extracted using a micropipette upon completing the exposure period. The volume and pH value are immediately measured to preserve sample integrity, ensuring accurate subsequent analyses. The gastric juice undergoes rigorous analysis, that includes free and total acidity determinations through titration using 0.01 N NaOH. The ulcer index is calculated based on observed outcomes, and the data are meticulously recorded. Added phenolphthalein indicator to a 1 mL sample, titrating with 0.01 N NaOH until a faint pink endpoint is achieved. The free acidity is calculated using the formula:

Free Acidity (mEq/L) = (Volume of NaOH (mL)times × Normality of NaOH)/(Volume of gastric juice sample (mL))

Given that the normality of NaOH is 0.01 N and the volume of the gastric juice sample is 1 mL, the formula simplifies to:

Free Acidity (mEq/L) = Volume of NaOH (mL) × 10

In one embodiment herein, the stomach is examined for the presence of ulcers. The ulcers are classified based on their severity into four categories: a score of 0 is assigned for no visible ulcers, 1 for superficial ulcers, 2 for deep ulcers, and 3 for perforated ulcers. This classification system allowed for the precise grading of ulcer severity during the examination.

The ulcer index is calculated using the following formula:

Ulcer Index= (Number of fishes with ulcers+ Total ulcer score)/(Total number of fishes)

After completing the titration for free acidity, Topfer’s reagent is added to the same sample. The titration continued until the color changed from red to yellowish-orange, indicating the endpoint. The total acidity is then calculated using the following formula:

Total Acidity(mEq/L) = (Volume of NaOH (V2) (mL)×Normality of NaOH×1000)/(Volume of gastric juice sample (mL))

In one embodiment herein, upon completion of the test protocols, the efficacy of the Kappaphycus-Vetiver powder formulation is assessed against the standard compound and control groups. Analytical data from pH value measurements, free and total acidity evaluations, along with other relevant indices, offer a thorough assessment of the formulation's performance, providing strong support for the advancement of this novel treatment.

At step 326, the primary composition 100 has phenomenal efficiency is concluded. In one embodiment herein, a formulation comprises Kappaphycus alvarezii and Vetiver, prepared using an advanced double-drying and multi-stage grinding process, demonstrated significant efficacy in treating ethanol-induced gastric ulcers in the aquatic species, such as Labeo rohita. The formulation is tested at three concentrations of 25%, 50%, and 75% and is found to promote complete ulcer resolution across all tested fish sizes, indicating its broad applicability.

In this embodiment, the ethanol is administered at varying concentrations of 1:3, 1:1, and 3:1 to induce gastric ulceration in the test subjects. The ulcers are systematically assessed using a standardized scoring system, where ulcerative lesions are classified as superficial, deep, or perforated. An ulcer index is calculated for each of the group based on the severity of the ulcers and the number of affected fish. The ulcer index revealed varying levels of gastric damage, providing a basis for evaluating the efficacy of the treatment.

Additionally, gastric juice samples are collected following the sacrifice of the test subjects to measure pH, free acidity, and total acidity. The analysis showed significant modulation of gastric acidity in the different groups. In the control group, baseline pH value and acidity levels are maintained, and no significant ulcer formation is observed. The Ranitidine group showed a moderate reduction in free and total acidity, which correlated with partial ulcer healing. Notably, the Kappaphycus alvarezii-Vetiver formulation exhibited a substantial reduction in both free and total acidity across all concentrations. This reduction in acidity levels contributed to the complete resolution of gastric ulcers in the treated fish.

In a comparative analysis of the ulcer index, the Kappaphycus alvarezii-Vetiver formulation demonstrated superior efficacy over the control and Ranitidine groups. In the control group, the ulcer index remained at zero due to the absence of ulceration. In the Ranitidine group, there is a moderate reduction in the ulcer index, indicating partial healing. However, in the test group treated with the Kappaphycus alvarezii-Vetiver formulation, the ulcer index is reduced to zero across all concentrations, signifying complete healing and superior performance compared to the Ranitidine group. Thus, this embodiment highlights the potential of the Kappaphycus alvarezii-Vetiver formulation as an effective natural treatment for ethanol-induced gastric ulcers in aquatic species, offering consistent therapeutic benefits across varying concentrations and fish sizes.

According to another example embodiment of the invention, FIG. 4 refers to a pictorial representation 400 of aquatic species with ethanol-induced gastric ulcers. The 30 fishes in three batches are exposed to the ethanol-induced ulcers at different concentrations of 3:1, 1:3, and 1:1. So the aquatic species are affected with the ulcer’s infection. The aquatic species are the Labeo rohita, carefully selected for uniformity in size, graded into different cohorts, ensuring consistent physiological parameters across all test groups. The ethanol solutions formulated in precise volumes corresponding to the required ulcerogenic intensity (1:3, 1:1, 3:1 ratios), ensuring controlled and repeatable results.

According to another example embodiment of the invention, FIG. 5 refers to the pictorial representation 500 of the aquatic species upon healing the ethanol-induced gastric ulcers. The primary composition 100 is made into different concentrations of 25%, 50% and 75% and is orally inserted to all the 30 fishes in three batches which had ulcers at different concentrations of 3:1, 1:3, and 1:1. They are observed for 7 days. Each of the batch consists of approximately 200 grams of dried Kappaphycus alvarezii and Vetiver, ground into a fine powder. The primary composition 100 prepared at 25%, 50%, and 75% concentrations, using 25 mL, 50 mL, and 75 mL of powder in 100 mL distilled water with the Carboxymethyl Cellulose (CMC) solution, respectively.

In one embodiment herein, the high-quality laboratory glassware (test tubes, beakers microsieves), micropipettes, calibrated measurement instruments (pH meter), tools for dissection and biological sample collection to ensure accurate and reproducible experimental conditions. Following the experimental period, all fish are sacrificed, and gastric juice is extracted with a micropipette for immediate volume and pH value measurement for both post ulcers and test powder insertion. Ulcers are induced using ethanol solutions, with ulcer formation carefully observed in three fish per cohort. Free and total acidity of the gastric juice for post ulcers and test powder insertion, are determined via titration with 0.01 N NaOH, ensuring precise results.

In one embodiment herein, the experimental setup requires standard laboratory power supply to operate equipment such as the pH meters and heating elements for the drying process of the materials. All experimental procedures are conducted under controlled temperature (approximately 25°C) conditions to ensure reproducibility and accuracy. Contamination free environment fosters the optimal working conditions for the experiment. Use of sterile instruments and precise handling techniques to prevent contamination and ensure accurate data collection. All biological waste, such as sacrificed fish and gastric contents, is disposed of following standard biosafety and environmental guidelines, ensuring no environmental contamination.

The Carboxymethyl Cellulose (CMC) solution ensures uniform suspension of the test compounds. The primary composition 100 effectively reduces both free and total gastric acidity, with the 75% concentration showing the highest efficacy. Ulcer Index measurements demonstrated a marked decrease in ulcer formation across all test concentrations. The formulation efficiently modulates gastric acidity with minimal energy inputs, making it cost-effective and scalable. The swift reduction in ulcer severity, particularly with the 75% concentration, highlights its rapid onset of action.

According to another example embodiment of the invention, FIG. 6 refers to a bar chart representation 600 of an efficiency of the primary composition 100. In one embodiment herein, the bar chart illustrates the total acidity levels under various test conditions, assessing the impact of different treatments on ethanol-induced gastric ulcers. The y-axis represents the ethanol concentration, while the x-axis presents three different treatment ratios of 3:1, 1:1, and 1:3. The chart depicts the response of various groups, such as a first group, a second group, a third group and a fourth group.

In one embodiment herein, the first group represents the control group, where no ethanol is applied, showing the baseline total acidity. The second group treated with ethanol to induce gastric ulcers, which demonstrates elevated acidity levels due to ethanol exposure. The third group treated with ranitidine, a conventional anti-ulcer medication, showing a reduction in total acidity compared to the ethanol group.

The fourth group treated with a combination of the carboxymethyl cellulose (CMC) and a blend of the Kappaphycus alvarezii and Vetiver (K+V), demonstrating a significant decrease in total acidity relative to the ethanol group. The fifth group treated solely with the Kappaphycus alvarezii and Vetiver (K+V) blend, further emphasizing its effectiveness in reducing total acidity in ethanol-induced gastric ulcers.

In the 3:1 ratio, the first group exhibits the highest acidity. The second group showing a reduction compared to the first group but still maintaining elevated levels. The third group exhibits a notable decrease in acidity. The fourth group is a CMC + K+V group and the fifth group is a K+V group, while the CMC + K+V and K+V groups show further reductions, with K+V group alone resulting in the lowest total acidity. The 1:1 ratio follows a similar pattern, where the K+V group significantly outperforms the other treatments in reducing acidity. In the 1:3 ratio, the trend continues, with the K+V group achieving the most substantial reduction in total acidity across all tested ratios.

This embodiment demonstrates that the novel composition containing Kappaphycus alvarezii and Vetiver effectively reduces total acidity in aquatic species with ethanol-induced gastric ulcers. The K+V blend consistently outperforms ranitidine and the other treatments, suggesting its potential as a natural remedy for treating gastric ulcers in aquatic species. The combination of CMC with K+V also shows efficacy, but K+V alone proves to be the most potent formulation across all tested ratios.

According to another example embodiment of the invention, FIG. 7 refers to a flowchart 700 of a method for preparing the primary composition 100. At step 702, the Kappaphycus alvarezii powder 102 and the Vetiver powder 104 are prepared through a dual-phase drying protocol followed by multi-stage grinding operation by using the Kappaphycus alvarezii material 10, Vetiveria zizanoides root’s material 12 and micro sieves 14. At step 704, the Kappaphycus alvarezii powder 102 and the Vetiver powder 104 are blended to obtain a homogeneous powder blend with optimal particle size distribution. At step 706, the homogeneous powder blend is dispersed in the distilled water at varying concentrations to prepare the primary composition 100 for precise dosing and application requirements.

Numerous advantages of the present disclosure may be apparent from the discussion above. In accordance with the present disclosure, the primary composition 100 and method for treating ulcers in aquatic species using Kappaphycus alvarezii and Vetiver powder, is disclosed. The proposed invention provides the primary composition 100 integrates Kappaphycus alvarezii and Vetiver powder, produced through a double-drying protocol and high-precision multi-stage grinding, to achieve an ultra-fine powder with exceptional uniformity and bioactivity. The cost-effective and sustainable method for preparing the Kappaphycus alvarezii and Vetiver powder formulation, requiring minimal resources and accessible to individuals with limited technical expertise. The method demonstrates the exceptional efficacy of the Kappaphycus alvarezii and Vetiver formulation in completely healing ethanol-induced gastric ulcers in fish, outperforming current treatment used in aquaculture.

The eco-friendly, chemical-free method for producing the formulation, enhancing ecological resilience and sustainability in aquaculture practices. The Kappaphycus alvarezii and Vetiver formulation underscores its broad-spectrum potential as a natural remedy for ulcers in aquaculture, effectively addressing common ailments worsened by environmental stressors. The primary composition 100 enhances palatability for fish, ensuring greater acceptance and easier administration compared to the other treatments, such as antibiotics, which carry the risk of promoting antibiotic resistance.

It will readily be apparent that numerous modifications and alterations can be made to the processes described in the foregoing examples without departing from the principles underlying the invention, and all such modifications and alterations are intended to be embraced by this application.
, Claims:CLAIMS:
I/We Claim:
1. A primary composition (100), comprising:
48 to 52 weight percentage of a Kappaphycus alvarezii powder (102) as an active ingredient, wherein the Kappaphycus alvarezii powder (102) is prepared through a dual-phase drying protocol followed by a multi-stage grinding operation; and
48 to 52 weight percentage of a Vetiver powder (104) as a natural ingredient, wherein the Vetiver powder (104) is prepared through the dual-phase drying protocol followed by the multi-stage grinding operation,
wherein the Kappaphycus alvarezii powder (102) and the Vetiver powder (104) are blended to obtain a homogeneous powder blend with optimal particle size distribution,
wherein the homogeneous powder blend is dispersed in distilled water at varying concentrations to prepare the primary composition (100) for precise dosing and application requirements,
wherein the primary composition (100) exhibits anti-ulcer activity.
2. The primary composition (100) as claimed in claim 1, wherein the Kappaphycus alvarezii powder (102) is derived from freshly harvested Kappaphycus alvarezii material (10), which is washed, sun-dried, and ground upon undergoing the dual-phase drying protocol.
3. The primary composition (100) as claimed in claim 1, wherein the Vetiver powder (104) is derived from Vetiveria zizanoides root’s material (12), which are washed, dried, and ground upon undergoing the dual-phase drying protocol.
4. The primary composition (100) as claimed in claim 1, wherein the Kappaphycus alvarezii powder (102) and Vetiver powder (104) are blended in at least a 1:1 ratio, selected based on empirical data demonstrating synergistic effects that enhance the performance of the primary composition (100).
5. The primary composition (100) as claimed in claim 1, wherein the dual-phase drying protocol includes an initial 24 hr natural solar drying phase followed by a controlled thermal drying phase at a temperature of 70 °C for a time period of 24 hr, thereby ensuring complete dehydration while preserving the structural integrity and active constituents of the Kappaphycus alvarezii powder (102) and the Vetiver powder (104).
6. The primary composition (100) as claimed in claim 1, wherein the Kappaphycus alvarezii powder (102) and the Vetiver powder (104) are obtained through a mechanical grinding and triple-stage micro-sieving process using micro sieves (14), thereby ensuring uniform particle size distribution and minimizing thermal degradation during the refinement process.
7. The primary composition (100) as claimed in claim 1, wherein the homogeneous powder blend is dispersed in the distilled water with carboxymethyl cellulose (CMC) as a vehicle at varying concentrations to prepare as a secondary composition (200) for precise dosing and application requirements,
wherein the homogeneous powder blend is dispersing in the distilled water with the carboxymethyl cellulose (CMC), which ensures stable dispersion of the active components, facilitating consistent and uniform interaction with aquatic subjects, such as the fish species (Labeo rohita).
8. A method for preparing a primary composition (100), comprising:
preparing a Kappaphycus alvarezii powder (102) and a Vetiver powder (104) through a dual-phase drying protocol followed by multi-stage grinding operation by using Kappaphycus alvarezii material (10), Vetiveria zizanoides root’s material (12) and micro sieves (14);
blending the Kappaphycus alvarezii powder (102) and the Vetiver powder (104) to obtain a homogeneous powder blend with optimal particle size distribution; and
dispersing the homogeneous powder blend in distilled water at varying concentrations to prepare the primary composition (100) for precise dosing and application requirements.