FIEED OF THE INVENTION

The present disclosure relates to drug discovery systems for medical-parasitic treatment, in more details, a system for producing anti-parasitic compounds and derivatives from marine hypotrichous ciliates.

BACKGROUND OF THE INVENTION

Ciliates are also used to test the toxicity of contaminants such detergents, heavy metals, carcinogenic chemicals, herbicides, insecticides, fungicides, antibiotics, and organic solvents in monoxenic cultures. Ciliates, on the other hand, have been utilized in holoxenic cultures to test the toxicity of heavy metals, herbicides, benzene, and phenol derivatives. The ability to determine the degree and possibility of ecological harm caused by anthropogenic pollution discharges to surface waters will be aided by understanding the ciliates' sensitivity to a wide range of hazardous compounds. Ciliates are now cultured in baseline solutions derived from freshwater, ocean, or vertebrate body fluids. These are supplemented with bacteria, fish cells, or organic substances, which can be specified or undefined, with proteose peptone being a good example.

In the view of the forgoing discussion, it is clearly portrayed that there is a need to have a system for producing anti-parasitic compounds and derivatives from marine hypotrichous ciliates.

SUMMARY OF THE INVENTION

The present disclosure seeks to provide a system for culturing protozoan hypotrichs anti-parasitic compounds and derivatives from marine hypotrichous ciliates.

In an embodiment, a system for producing anti-parasitic compounds and derivatives from marine hypotrichous ciliates is disclosed. The system includes a centrifuge for centrifuging 200-300mL sieved faeces extract slurry at 30000rpm for 20-40mins at room temperature. The system further includes a treating unit for filtering or sterilizing the supernatant fraction. The system further includes a main flask for mixing 4-6g of fresh equine faeces and 30-40mL of the respective mineral salt solutions, wherein the cultures are fed every day and saturated with CO2 for 5-10min at a flow velocity of 60 cm gas min . The system further includes an agitator for receiving the 15-30g fresh culture into fresh medium after a predetermined time for agitating it before adding 15-30mL aliquots of existing culture medium. The system further includes a water bath assembly for performing all manipulations at 40°C in the presence of CO2 thereby storing in an inoculated flask and sealing with a rubber stopper, wherein the inoculated flask is further stored in an incubator at 40°C.

In an embodiment, the largest plant particles are removed from extract using a sieve.

In an embodiment, the food is added to the inoculated flask, wherein the food is supplied at the level of 0.10-0.30 mg mL” ¹ per day.

In an embodiment, the cultures are transferred into fresh medium every fourth day, wherein the transferring includes adding preferably 20mL of pre-heated fresh culture medium to the main flask and agitating before adding preferably 20 mL aliquots of existing culture medium.

In an embodiment, the system comprises a scrapper unit for extracting 400-600 mL faeces extract from freshly faeces and caudatum salt solution and mixing in 1:2 ratio thereby removing largest plant particles.

In an embodiment, the treating unit comprises: a bacterial filter for collecting and filtering the obtained supernatant fraction; and a steriliser apparatus for sterilizing the supernatant fraction at 100°C for 20-30mins.

An object of the present disclosure is to culture protozoan ciliates.

Another object of the present disclosure is to develop Anti-Parasitic Compounds and Derivatives from Marine Hypotrichous Ciliates. Another object of the present disclosure is to make more efficiency drugs to the present parasites without adverse effect.

Yet another object of the present invention is to maintenance of protozoan ciliate culture.

To further clarify advantages and features of the present disclosure, a more particular description of the invention will be rendered by reference to specific embodiments thereof, which is illustrated in the appended drawings. It is appreciated that these drawings depict only typical embodiments of the invention and are therefore not to be considered limiting of its scope. The invention will be described and explained with additional specificity and detail with the accompanying drawings.

BRIEF DESCRIPTION OF FIGURES

These and other features, aspects, and advantages of the present disclosure will become better understood when the following detailed description is read with reference to the accompanying drawings in which like characters represent like parts throughout the drawings, wherein:

Figure 1 illustrates a block diagram of a system for producing anti-parasitic compounds and derivatives from marine hypotrichous ciliates in accordance with an embodiment of the present disclosure;

Figure 2 illustrates Table 1 depicts behavioral parameters test on test sample in accordance with an embodiment of the present disclosure; and

Figure 3 illustrates Table 2 depicts anti-inflammatory activity on test sample against Carrageenan induced Paw Edema in Albino Rat in accordance with an embodiment of the present disclosure.

Further, skilled artisans will appreciate that elements in the drawings are illustrated for simplicity and may not have necessarily been drawn to scale. For example, the flow charts illustrate the method in terms of the most prominent steps involved to help to improve understanding of aspects of the present disclosure. Furthermore, in terms of the construction of the device, one or more components of the device may have been represented in the drawings by conventional symbols, and the drawings may show only those specific details that are pertinent to understanding the embodiments of the present disclosure so as not to obscure the drawings with details that will be readily apparent to those of ordinary skill in the art having benefit of the description herein.

DETAILED DESCRIPTION:

For the purpose of promoting an understanding of the principles of the invention, reference will now be made to the embodiment illustrated in the drawings and specific language will be used to describe the same. It will nevertheless be understood that no limitation of the scope of the invention is thereby intended, such alterations and further modifications in the illustrated system, and such further applications of the principles of the invention as illustrated therein being contemplated as would normally occur to one skilled in the art to which the invention relates.

It will be understood by those skilled in the art that the foregoing general description and the following detailed description are exemplary and explanatory of the invention and are not intended to be restrictive thereof.

Reference throughout this specification to “an aspect”, “another aspect” or similar language means that a particular feature, structure, or characteristic described in connection with the embodiment is included in at least one embodiment of the present disclosure. Thus, appearances of the phrase “in an embodiment”, “in another embodiment” and similar language throughout this specification may, but do not necessarily, all refer to the same embodiment.

The terms "comprises", "comprising", or any other variations thereof, are intended to cover a non-exclusive inclusion, such that a process or method that comprises a list of steps does not include only those steps but may include other steps not expressly listed or inherent to such process or method. Similarly, one or more devices or sub-systems or elements or structures or components proceeded by "comprises...a" does not, without more constraints, preclude the existence of other devices or other sub-systems or other elements or other structures or other components or additional devices or additional sub-systems or additional elements or additional structures or additional components. Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. The system, methods, and examples provided herein are illustrative only and not intended to be limiting.

Embodiments of the present disclosure will be described below in detail with reference to the accompanying drawings.

Referring to Figure 1, a block diagram of a system for producing anti -parasitic compounds and derivatives from marine hypotrichous ciliates is illustrated in accordance with an embodiment of the present disclosure. The system 100 includes a scrapper unit 102 for extracting 400-600 mL faeces extract from freshly faeces and caudatum salt solution and mixing in 1:2 ratio thereby removing largest plant particles.

In an embodiment, a centrifuge 104 is connected to the scrapper unit 102 for centrifuging 200-300mL sieved faeces extract slurry at 30000rpm for 20-40mins at room temperature.

In an embodiment, a treating unit 106 is connected to the centrifuge 104 for filtering or sterilizing the supernatant fraction.

In an embodiment, a main flask 108 is connected to the treating unit 106 for mixing 4- 6g of fresh equine faeces and 30-40mL of the respective mineral salt solutions, wherein the cultures are fed every day and saturated with CO2 for 5-10min at a flow velocity of 60 cm ³ gas min ^-1 .

In an embodiment, an agitator 110 is connected to the main flask 108 for receiving the 15-30g fresh culture into fresh medium after a predetermined time for agitating it before adding 15-30mL aliquots of existing culture medium.

In an embodiment, a water bath assembly 112 is connected to the agitator 110 for performing all manipulations at 40°C in the presence of CO2 thereby storing in an inoculated flask and sealing with a rubber stopper, wherein the inoculated flask is further stored in an incubator 114 at 40°C.

In an embodiment, the largest plant particles are removed from extract using a sieve.

In an embodiment, the food is added to the inoculated flask, wherein the food is supplied at the level of 0.10-0.30 mg mL” ¹ per day.

In an embodiment, the cultures are transferred into fresh medium every fourth day, wherein the transferring includes adding preferably 20mL of pre-heated fresh culture medium to the main flask 108 and agitating before adding preferably 20 mL aliquots of existing culture medium.

In an embodiment, the treating unit 106 comprises a bacterial filter 116 for collecting and filtering the obtained supernatant fraction. The treating unit 106 further comprises a steriliser apparatus 118 for sterilizing the supernatant fraction at 100°C for 20-30mins.

Protozoan Parasitic laboratory culture is very tedious nature rather than plating of bacteria and viruses. If we culture in the laboratory procedures, the laboratory is highly contaminated and infected. This is first disadvantages. The second is media selection procedure complicated and always used in special media procedures. And third risk of maintenance of protozoan ciliate culture. More or less all the synthetic drugs have more adverse effects.

Figure 2 illustrates Table 1 depicts behavioral parameters test on test sample in accordance with an embodiment of the present disclosure.

Figure 3 illustrates Table 2 depicts anti-inflammatory activity on test sample against Carrageenan induced Paw Edema in Albino Rat in accordance with an embodiment of the present disclosure.

About the Species:

Euplotes cristatus often habitat in estuary water of Rameshwaram areas, particularly in Tirupulani and Kilakarai coastal areas of Tamilnadu. Salinity, temperature and other parameters are important factors for sustain this species. The researcher learnt, if BOD is less in water, and DO has increased, the Euplotes cristatus and other related strains more abundance in the water observed. But if BOD is increased, the researcher has taken less microbial content in the water. Euplotes spp. have 55micron to 65 microns in long and often oval in shaped. The Adoral Zone of Membranelles (AZM) always in curve shaped 35 to 45 membranelles were observed. The dorsal argyrome is often 8 dorso-lateral kinetics and 11 to 15 dorsal cilia in the central rows. It is key features for identification of species. It is very unique for the species as per the report of Kahl (1932) identification procedures. Micronucleus and macronucleus are prominently identified for this species.

Industrial Benefits:

This compounds and their derivatives of Marine hypotrichs, control the growth of human parasites of Entamoeba histolytica sp. This compound and derivatives secreted for control of intra and inter species of other varieties in nature. It is little effect on the prokaryotic groups. At present, we use the synthetics drug of anti-parasitic combination like albendazole more adverse effects such as nausea, vomiting, hair-loss and abdominal pain. But this nature compound and derivatives the researcher verified in-vivo test in white albino rat. The in-vitro and in-vivo test verified in Periyar College of Pharmaceutical Sciences, Tiruchirappalli with ethical clearance certificate. Other derivatives verified in the Govt. KAP Vishvanatham Medical College Tiruchirappalli with Institutional Ethics Committee from Directorate of Medical Education, Chennai. No such effects observed during the research study.

Culture Techniques:

While culturing this species in the controlled laboratory procedure, the researcher faced variety of aseptic maintenance. Bacteria can grow in the artificial media. But protozoan parasitic culture is difficult one. With the help of doctors and laboratory technicians, the researcher got the pure culture of parasite and successfully verified.

The developed system 100 produces inter-specific relationship control product from the organisms. It is purely secondary metabolites. Hence, no side effects observed from this discovery invitro and in-vivo. The anti-parasitic compound is prepared by isolation of hypotrichs from marine waters (Estuary). The pure culture is taken from the mixed population. The compound is evaluated by column chromatography techniques, HPLC and NMR techniques to identify the compound. The compound showed anti-microbial activity.

In-vitro and in-vivo techniques is observed with two ethical clearance certificates.

The drawings and the forgoing description give examples of embodiments. Those skilled in the art will appreciate that one or more of the described elements may well be combined into a single functional element. Alternatively, certain elements may be split into multiple functional elements. Elements from one embodiment may be added to another embodiment. For example, orders of processes described herein may be changed and are not limited to the manner described herein. Moreover, the actions of any flow diagram need not be implemented in the order shown; nor do all of the acts necessarily need to be performed. Also, those acts that are not dependent on other acts may be performed in parallel with the other acts. The scope of embodiments is by no means limited by these specific examples. Numerous variations, whether explicitly given in the specification or not, such as differences in structure, dimension, and use of material, are possible. The scope of embodiments is at least as broad as given by the following claims.

Benefits, other advantages, and solutions to problems have been described above with regard to specific embodiments. However, the benefits, advantages, solutions to problems, and any component(s) that may cause any benefit, advantage, or solution to occur or become more pronounced are not to be construed as a critical, required, or essential feature or component of any or all the claims.