ANTIBACTERIAL AND ANTIMYCOBACTERIAL PROPERTY OF CRUDE EXTRACT OF BOX JELLY FISH CHIROPSOIDES BUITENDIJKI (HORST R, 1907) FROM WEST COAST OF MUMBAI

ANTIBACTERIAL AND ANTIMYCOBACTERIAL PROPERTY OF CRUDE EXTRACT OF BOX JELLY FISH CHIROPSOIDES BUITENDIJKI (HORST R, 1907) FROM WEST COAST OF MUMBAI

J. L. Dolnar and G. V. Zodape *

Department of Zoology, S. S. & L.S. Patkar College of Arts & Science & V. P. Varde College of Commerce & Economics S. V. Road Goregaon (west), Mumbai, Maharashtra, India.

ABSTRACT: The Box jelly fish Chiropsoides buitendijki (Horst R, 1907) were collected during low tides from Edwan village, of West Coast of Mumbai. Identification was done at the Central Marine Fisheries Research Institute (CMFRI), Mumbai. Crude extract of Chiropsoides buitendijki (Horst R, 1907) was obtained following the 80% methanol and 1%acetic acid, absolute methanol and n-butanol (1:1), and n-butanol. The antibacterial activity was carried out on bacteria E. coli, K. pneumonia, S. aureus, S. flexneri, V. Cholerae, C. sporogenes, S. enterica, E. faecalis, and fungal strain Candida albicans and Aspergillus niger whereas the antimycobacterial study was carried out on Mycobacterium tuberculosis (MTB) strain, H37 Rv: ATCC No- 27294. From the above study it is confirms that, the crude extract of Chiropsoides buitendijki have antibacterial and anti- tuberculosis property.

Keywords: Jelly fish, Edwan village, Crude extract, Antibacterial, Antimycobacterial property

INTRODUCTION: Many studies investigated bioactive compounds from marine animals for various purposes including anti-bacterial, anti-tumor, anti-inflammatory, nutritional supplements, discovery of novel drugs and its applications for industrial biotechnology ¹. Several marine bioactive compounds are of great benefit to pharmaceutical industries to prepare human medicines including cancer therapy, vascular diseases and infectious viral diseases such as AIDS ². Jelly fish belongs to a group of free-swimming animals that belongs to the phylum Cnidaria. Cnidarians have special cells called cnidocytes which gives the name to the phylum Cnidarians.

The world fishery for edible jellyfish, which is mostly based in Southeast Asia, produces an annual catch of more than 750,000 tones, with rising demand reaching outside of Asian markets. Jellyfish appear to have qualities of a nutritious diet and are known for their nutritional and medicinal significance in the Chinese pharmacopoeia. Certain Cnidaria jellyfish species from the Mediterranean can serve as an affordable source of raw materials for cutting-edge pharmaceutical, nutraceutical, or cosmetic goods.

Because of their high collagen content and protein content, jellyfish have just been recognized in Europe as novel foods. Jellyfish proteins and other isolated substances have significant antioxidant and other biological properties. Many studies are found on cnidocytes for several purposes, but very scant study is available on their toxic potential ³. However, neurotoxin is found in the ectodermal gland cells ⁴.

Therefore, extensive study is required not only on the nematocyst but also the whole body of jelly fish ⁵. Jellyfish stings can cause a burning sensation, excruciating pain, swelling, red streaks, nausea, cramps in the stomach, excessive perspiration, respiratory trouble, heart failure, and other symptoms ⁶. In line with this, jellyfish venoms exhibit a diverse range of biological effects, including dermonecrotic, neurotoxic, hemolytic, and cardiovascular effects ^{7, 8}. Acute heart failure is acknowledged as the primary cause of mortality brought on by jellyfish venoms, and it is thought that the effects of jellyfish venoms are brought on by the combination of numerous poisonous components ^{9, 10}. Countless number of people is found to be poisoned by jelly fish every year throughout the world, but statistical data is rarely available, even though the fatal cases are not available in this regard. Many research showed the regime for the treatment against cnidarian stings, but confusion still exists as to what is the most effective first aid and clinical management against cnidarians toxin. Significant research is available for the treatment on cnidarian stings, but only a few toxic components have been identified so far, it is one of the most neglected areas in toxicology research. This is most likely due to the difficulty in obtaining jellyfish, which, in contrast to other venomous invertebrates, are widely dispersed in the open ocean. Second, unlike the milking of snake venom, it is very challenging to gather high-purity jellyfish venom without contaminating it with other tissue detritus. Lastly, because all of the poisons discovered from jellyfish venoms up to this point are proteins, many of which are innately vulnerable to harsh environmental conditions and readily denatured, it is necessary to identify the dangerous components found in jellyfish.

MATERIALS AND METHODS:

Collection of Samples: The Box jelly fish Chiropsoides buitendijki (Horst, R 1907) were collected during low tides from Edwan village, of West Coast of Mumbai. Animals were taken alive to the laboratory in sea water washed under sea water and then with distilled water.

Identification of Box Jelly Fish: Preliminary identification was done by studying the shape and no. of tentacles and by referring the relevant literature and final confirmation of identification was done by Dr. Ramkumar, scientist, at the Central Marine Fisheries Research Institute (CMFRI), Mumbai.

Preparation of Box Jelly Fish Crude Extracts: Crude extract of Chiropsoides buitendijki (Horst, R 1907)  was obtained following the 80% methanol and 1% acetic acid, absolute methanol and n-butanol (1:1), and n-butanol by applying method of ¹¹ with some modifications. 10 grams of Box jelly fish samples; 10 ml mixture of equal volume of 80 % methanol and 1%acetic acid, 10 ml of absolute methanol and n-butanol (1:1) and 10 ml of n-butanolwas added and kept standing for 24 hrs. Solvent were then removed, by squeezing Box jelly fish samples, and filtered through Whatman filter paper No.1. The homogenate centrifuged at 5000 rpm for 15 minutes in cold centrifuge at -8°C and the supernatant was collected. The remaining solvent was evaporated at low pressure using Rotary Vacuum Evaporator at 45⁰C. The resultant compound was subjected to Millipore filter system and finally dried in vacuum desiccators and stored at 4⁰C in a refrigerator till further use for antibacterial and antimycobacterial study.

Ethical Approval: Ethical approval is sought from Maharashtra State Biodiversity Board, Nagpur, Maharashtra for collection of Box jelly fish samples for research purpose (No.:MSBB/Desk-5/ /Research/ 841/2022-23).

Procurement of Bacterial Cultures: The pure culture of bacteria E. coli, K. pneumoniae, S.aureus, S. flexneri, V. cholerae, C. sporogenes, S. enterica, E. faecalis and fungal strain Candida albicans and Aspergillus niger were collected from APX laboratories, Plot No. B7/1, Kothari Warehouse No.3, S. V. Road, Chitalsar, Manpada, Thane west-607, Mumbai, India and Mycobacterium tuberculosis (MTB) strain, H37 Rv: ATCC No- 27294 was procured from Maratha Mandal's Central Research Laboratory, Maratha Mandal's NGH Institute of Dental Sciences and Research Centre, R.S.No. 47A/2, Bauxite Road, Belgaum-590010, India.

Antibacterial and Antimycobacterial Study: The antibacterial study was carried out by using bacteria seeded plate –Well method as proposed by ¹² and the anti-Mycobacterial activity of crude extracts of Chiropsoides buitendijki (Horst, R 1907)   were assessed against M. tuberculosis strain H37 Rv: ATCC No- 27294 using microplate alamar blue assay (MABA) as proposed by ¹³.

RESULTS AND DISCUSSION:

TABLE 1: SHOWING EFFECT OF CRUDE EXTRACT OF CHIROPSOIDES BUITENDIJKI (HORST, R 1907) IN DIFFERENT ORGANIC MIXTURES ON DIFFERENT BACTERIAL AND FUNGAL STRAINS IS IN THE FORM OF ZONE OF INHIBITION

TABLE 2: SHOWING EFFECT OF CRUDE EXTRACT OF CHIROPSOIDES BUITENDIJKI (HORST, R1907) IN DIFFERENT ORGANIC MIXTURES AND STANDARD DRUGS ON M. TUBERCULOSIS STRAIN H37 RV: ATCC NO- 27294 USING MICROPLATE ALAMAR BLUE ASSAY (MABA)

* S= Sensitive *R=Resistant

PHOTOGRAPH 1: PHOTOGRAPH SHOWING, A) EFFECT OF CRUDE EXTRACT OF CHIROPSOIDES BUITENDIJKIIN DIFFERENT ORGANIC MIXTURES B) STANDARD DRUG CONCENTRATIONS ON M. TUBERCULOSIS STRAIN H37 RV: ATCC

DISCUSSION: Box jelly fish Chiropsoides buitendijki (Horst, R1907) belonging to the phylum cnidaria and class cubozoa have been reported for its multitudinous medicinal values. The study carried out by ¹⁴ showed antibacterial activity against toxin of Anemonia sulcata. In another study ¹⁵ have isolated the neurotoxin Ueq 12-1 from Anemonia sulcata and Urticina eques showed Corynebacterium glutamicum, and Staphylococcus aureus bacterial inhibition of human pathogens ¹⁶. studied on cytolytic actinoprins, cardio stimulatory proteins and cytolysins isolated from sea anemones; Heteractis magnifica and Stichodactyla mertensii showed higher antibacterial potentials against Staphylococcus aureus and Salmonella typhi. According to ¹⁷ sea anemone neurotoxin was found effective against Bacillus subtilis at minimal effective concentration and showed moderate antibacterial potency against E. coli and Salmonella enteric ¹⁸. studied the methanolic extract of nematocysts of Stichodactyla mertensii and Stichodactyla gigantean showed antibacterial property against Staphylococcus aureus, Salmonella typhi and Vibrio cholera ¹⁹. Isolated the bioactive compounds of Leptogorgia virgulata of the family Gorgoniidae showed antibacterial property. According to ²⁰ the active component homarine and a homarine analog showed inhibited growth of Vibrio harveyii and Micrococcus luteus, indicating antibacterial activity ²¹. Isolated mesoglea of jellyfish Aurelia aurita exhibited higher activity against Gram- negative and Gram-positive bacteria further ²². Isolated the venom from jelly fish Chrysaora quinquecirrha found moderately effective on human pathogens (Escherichia coli, Vibrio cholerae, Salmonella paratyphi, Klebsiella pneumoniae, Pseudomonas aeruginosa, Proteus mirabilis, Proteus vulgaris and Klebsiella oxytoca), whereas Salmonella paratyphi is found more sensitive against venom of Chrysaora quinquecirrha.

The present study was undertaken to examine the effect of crude extract of Chiropsoides buitendijki prepared in different organic solvents, 80% methanol and 1%acetic acid, absolute methanol and n-butanol (1:1), and n-butanol to find antibacterial property.  The crude extract of Chiropsoides buitendijki was also tested on standard drugs Isoniazid, Ethambutol, Pyrazinamide, Rifampicin, and Streptomycin against Mycobacterium tuberculosis (MTB) strain, H37 Rv: ATCC No- 27294 to find the antimycobacterial property. From Table 1 Showing effect of crude extract of Chiropsoides buitendijki (Horst, R 1907) in different organic mixtures on different bacterial and fungal strains is in the form of zone of inhibition. It was observed that in 80 % methanol and 1%acetic acid against all the bacterial and fungal strainsno zone of inhibition was observed. In case of absolute methanol and n-butanol (1:1), the zones of inhibitions were observed against S. flexneri (8.16mm),V. cholerae (19.44mm), C. sporogenes (20.17mm), S. enterica (36.19mm), and E. faecalis (14.31mm) respectively. In n-butanol the zone of inhibition was observed in E. coli (8.3mm), K. pneumoniae (9.0mm), S. aureus (9.83mm), S. flexneri (9.16mm), V. cholerae (19.34mm), C. sporogenes (20.26mm), S. enterica (40.03mm), and E. faecalis (25.23mm) respectively. In case of fungal strains Candida albicans and Aspergillus niger no zone of inhibitions was noted in different organic mixtures. Table 2 Showing effect of crude extract of Chiropsoides buitendijki in different organic mixtures and standard drugs on M. tuberculosis strain H37 Rv: ATCC No- 27294 using microplate alamar blue assay (MABA) and Photograph: 1, Photograph Showing, a) Effect of crude extract of Chiropsoides buitendijkiin different organic mixtures b) standard drugs against Mycobacterium tuberculosis (MTB) strain H37 Rv: ATCC No- 27294.

The sensitivity of the crude extract and standard drugs against Mycobacterium tuberculosis (MTB) strain, H37 Rv: ATCC No- 27294 and it was found that in Isoniazid (1.6 µg/ml), Ethambutol (1.6 µg/ml), Pyrazinamide (3.125µg/ml), Rifampicin (0.8µg/ml), and Streptomycin (0.8µg/ml) respectively. The sensitivity of crude extract prepared in different organic solvents was   also checked against Mycobacterium tuberculosis (MTB) strain, H37 Rv: ATCC No- 27294 and it was found that in 80 % methanol and 1%acetic acid (12.5µg/ml), in absolute methanol and n-butanol (1:1) (50µg/ml), and in n-butanol (50µg/ml) respectively.

From the above results it is confirmed that Box jelly fish Chiropsoides buitendijki (Horst, R1907) contains bioactive compounds which showed antibacterial and antimycobacterial property.

CONCLUSION: Marine animals produce verities of natural toxins in order to utilize them for their defense mechanism and to protect them from other predators. Recently, many marine toxins have risen in the field of novel drug discovery and proven its wide spectrum of pharmaceutical and biomedical potential. However, with respect to the Cnidarian toxins the efforts have reveals on its neurotoxins potentials that are being used for various therapeutic applications in nervous system and very scanty reports are available on its bioactive compounds. The screening of Cnidarian bioactive compounds and evaluating its activity on different animal model is highly recommended and there is a need to be a major focus in drug discovery and therapy. Therefore, we have screened the effect of crude extract of Box jelly fish Chiropsoides buitendijki (Horst, R1907) ant it confirms that the Box jelly fish contains bioactive compounds which showed antibacterial and antimycobacterial property.

ACKNOWLEDGEMENT: Authors are thankful to Dr. Ramkumar, scientist, at the Central Marine Fisheries Research Institute (CMFRI), Mumbai for final identification and confirmation of species. Authors are also thankful to the Director, Maharashtra State Biodiversity Board, Nagpur, Maharashtra for giving permission for collection of species. Thanks are also due to APX laboratories, Thane, Maharashtra for providing bacterial strains and, Maratha Mandal's Central Research Laboratory, Belgaum-590010, India, for Mycobacterium tuberculosis (MTB) strain H37 Rv: ATCC No- 27294.

CONFLICT OF INTEREST: Authors have no conflict of interest

REFERENCES:

1. Urbarova I, Karlsen B, Okkenhaug S, Seternes O, Johansen S and Emblem A: Digital marine bioprospe cting: Mining new neurotoxin drug candidates from the transcriptomes of cold-water sea anemones. Mar Drugs 2012; 10: 2265–2279.
2. Wei WC, Sung P, Duh C, Chen B, Sheu J and Yang NS: Anti-inflammatory activities of natural products isolated from soft corals of Taiwan between 2008 and 2012. Mar. Drugs 2013; 11: 4083–4126.
3. Moran Y, Genikhovich G, Gordon D, Wienkoop S, Zenkert C, Ozbek S, Technau U and Gurevitz M: Neurotoxin localization to ectodermal gland cells uncovers an alternative mechanism of venom delivery in sea anemones. Proc Biol Sci 2012; 279: 1351–1358.
4. Frazao B, Vasconcelos V and Antunes A: Sea anemone (Cnidaria, Anthozoa, Actiniaria) toxins: An overview. Mar. Drugs 2012; 10: 1812–1851.
5. Frazao B, Vasconcelos V and Antunes A: Sea anemone (Cnidaria, Anthozoa, Actiniaria) toxins: An overview. Mar. Drugs 2012; 10: 1812–1851.
6. Suganthi K, Bragadeeswaran S, Sri Kumaran N, Thangaraj S and Balasubramanian T: Biological and pharmacological activities of jelly fish Crambionella stuhalmanni (Chun, 1896) and Chrysaora quinquecirrha (Desor, 1843). Inter J Pharm. harmaceut Sci 2011; 3(2): 230-236.
7. Tibballs J: Australian venomous jellyfish, envenomation syndromes, toxins and therapy, Toxicon 2006; 48: 830-859.
8. Suput D: In-vivo effects of cnidarian toxins and venoms. Toxicon 2009; 54: 1190-1200.
9. Currie B: Clinical implications of research on the box-jellyfish Chironex fleckeri, Toxicon 1994; 34: 1305-1313.
10. Ramasamy S, Isbister GK, Seymour JE and Hodgson WC: The in-vivo cardiovascular effects of box jellyfish Chironex fleckeri venom in rats: efficacy of pre-treatment with antivenom, verapamil and magnesium sulphate. Toxicon 2004; 43: 685-690.
11. Braekman JC, Daloze D, Stoller C and van Soest RWM: Chemotaxonomy of Agelas (Porifera: Demospongiae). Biochem. Syst Ecol 1992; 20: 417–431.
12. Heatley NG: A method for the assay of penicillin. Biochem J 1944; 38: 61–65.
13. Maria CS, Lourenço, Marcus V. N. de Souza, Alessandra C. Pinheiro, Marcelle de L. Ferreira, Raoni S, Gonçalves, Thais Cristina M. Nogueira and Mônica A. Peraltab: Evaluation of anti-tubercular activity of nicotinic and isoniazid analogues. ARKIVOC 2007; 181-191.
14. Trapani MR, Parisi MG, Toubiana M, Coquet L, Jouenne T, Roch P and Cammarata M: First evidence of antimicrobial activity of neurotoxin 2 from Anemonia sulcata (Cnidaria). Invertebrate Survival Journal 2014; 11(1): 182–191.
15. Logashina YA, Solstad RG, Mineev KS, Korolkova YV, Mosharova IV, Dyachenko IA, Palikov VA, Palikova YA, Murashev AN and Arseniev AS: New disulfide-stabilized fold provides sea anemone peptide to exhibit both antimicrobial and TRPA1 potentiating properties. Toxins (Basel) 2017; 9(5): 154.
16. Ghosh S, Ajith Kumar TT and Balasubramanian T: Characterization and antimicrobial properties from the sea-anemones (Heteractics magnifica and Stichodactyla mertensii) toxins. Archives of Applied Science Research 2011; 3(4): 109–117.
17. Kim CH, Lee YJ, Go HJ, Oh HY, Lee TK, Park JB and Park NG: Defensin-neurotoxin dyad in a basally branching metazoan sea anemone. The FEBS Journal 2017; 284(19): 3320–3338.
18. Thangaraj S, Bragadeeswaran S, Suganthi K and Sri Kumaran N: Antimicrobial properties of sea anemone Stichodactyla mertensii and Stichodactyla gigantea from Mandapam coast of India. Asian Pacific Journal of Tropical Biomedicine 2011; 1(1): 43–6.
19. Shapo JL, Moeller PD and Galloway SB: Antimicrobial activity in the common sea whip, Leptogorgia virgulata (Cnidaria: Gorgonaceae). Comparative Biochemistry and Physiology B 2007; 148(1): 65–73.
20. Sun P, Meng LY, Tang H, Liu BS, Li L, Yi Y and Zhang W: Sinularosides A and B, bioactive 9,11-secosteroidal glycosides from the South China Sea soft coral Sinularia humilis Ofwegen. Journal of Natural Products 2012; 75(9): 1656–1659.
21. Ovchinnikova TV, Balandin SV, Aleshina GM, Tagaev AA, Leonova YF, Krasnodembsky ED, Men’shenin AV and Kokryakov VN: Aurelin, a novel antimicrobial peptide from jellyfish Aurelia aurita with structural features of defensins and channel-blocking toxins. Biochemical and Biophysical Research Communications 2006; 348(2): 514–523.
22. Suganthi K, Bragadeeswaran S: Antimicrobial and Immunomodulatory activities of Jellyfish (Chrysaora quinquecirrha) venom. In: Sabu A, Augustine A, editors, Prospects in bioscience: Addressing the issues. India Springer 2012; 283–292.
    

    ---

    How to cite this article:

    Dolnar JL and Zodape GV: Antibacterial and antimycobacterial property of crude extract of box jelly fish Chiropsoides buitendijki (horst r, 1907) from west coast of Mumbai. Int J Pharm Sci & Res 2023; 14(10): 4918-23. doi: 10.13040/IJPSR.0975-8232.14(10).4918-23.

    ---

    All © 2023 are reserved by International Journal of Pharmaceutical Sciences and Research. This Journal licensed under a Creative Commons Attribution-NonCommercial-ShareAlike 3.0 Unported License.