Brine Shrimp Larvae Research Articles

Comparative evaluation of impact of Zn and ZnO nanoparticles on brine shrimp (Artemia salina) larvae: effects of particle size and solubility on toxicity.

Brine shrimp (Artemia salina) larvae were exposed to different sizes of zinc (Zn) and zinc oxide (ZnO) nanoparticles (NPs) to evaluate their toxicity in marine aquatic ecosystems. Acute exposure was conducted in seawater with 10, 50 and 100 mg L(-1) concentrations of the NPs for 24 h and 96 h. Phase contrast microscope images confirmed the accumulation of the NPs inside the guts. Artemia were unable to eliminate the ingested particles, which was thought to be due to the formation of massive particles in the guts. Although the suspensions of the NPs did not exhibit any significant acute toxicity within 24 h, mortalities increased remarkably in 96 h and escalated with increasing concentration of NP suspension to 42% for Zn NPs (40-60 nm) (LC50∼ 100 mg L(-1)) and to about 34% for ZnO NPs (10-30 nm) (LC50 > 100 mg L(-1)). The suspensions of Zn NPs were more toxic to Artemia than those of ZnO NPs under comparable regimes. This effect was attributed to higher Zn(2+) levels (ca. up to 8.9 mg L(-1)) released to the medium from Zn NPs in comparison to that measured in the suspensions of ZnO NPs (ca. 5.5 mg L(-1)). In addition, the size of the nanopowders appeared to contribute to the observed toxicities. Although the suspensions possessed aggregates of comparable sizes, smaller Zn NPs (40-60 nm) were relatively more toxic than larger Zn NPs (80-100 nm). Likewise, the suspensions of 10-30 nm ZnO NPs caused higher toxicity than those of 200 nm ZnO NPs. Lipid peroxidation levels were substantially higher in 96 h (p < 0.05), indicating that the toxic effects were due to the oxidative stress.

In vitro antimicrobial and cytotoxic activity of Tamarix dioica Roxb. leaves

Medicinal plants have been in use for thousands of years for treatment of different types of diseases and a large inventory of useful drugs has been isolated from them. The aim of our study is to investigate a new and effective plant possessing antimicrobial activity. The crude extracts of Tamarix dioica Roxb. leaves, as well as fractions of the extract in various solvents, were investigated against standard strains of bacteria Pseudomonas aeruginosa, Escherichia coli, Klebsiella pneumoniae, Shigella flexneri, Bacillus subtilis, Salmonella typhi, and Staphylococcus aureus and fungi Candida glabrata, Aspergillus niger, Trichophyton rubrum, Candida tropicalis, and Aspergillus fumigatus. For in vitro antibacterial and antifungal studies of T. dioica extracts, the agar well diffusion technique was used. For cytotoxic study, brine shrimp (Artemia salina) larvae were used. Results of the crude extract and some fractionated samples showed significant antifungal properties, but low antibacterial response and negligible cytotoxic activity. It was recognized that Tamarix dioica has the potential for future development of new antifungal drugs/medicine.

Antioxidant capacity versus chemical safety of wheat bread enriched with pomegranate peel powder

Pomegranate peel powder (PP), a by-product of the pomegranate juice industry rich in polyphenols, was explored for use in bread production, due to its potential health effects. Wheat bread was prepared using different levels for replacement of flour with PP (0 to 10 g per 100 g flour) resulting in antioxidant levels expressed as Trolox equivalent antioxidant capacity values (TEAC) ranging from 1.8 to 6.8 μmol TEAC per g bread for fresh bread. TEAC remained constant during 5 days of storage in polyethylene bags at room temperature. The oxidative stability was evaluated by detection of radicals by direct electron spin resonance (ESR) spectroscopy, and peroxide value, and the highest capacity of scavenging of radicals (Fremy's salt) and the lowest content of peroxide values were found in bread with the highest percentage of PP. Safety evaluation was performed by the Artemia salina assay. An increased death rate of the brine shrimp larvae was found as a function of the replacement of wheat flour with PP in fortified bread providing a general screening method for the toxicological test of polyphenol fortified bread to be recommended for use in product development in addition to subjective evaluation. Based on both toxicological and subjective evaluations an addition of 2.5% PP is recommended for the actual product.

Biological and chemical diversity of cytotoxin-producing symbiotic marine fungi in intertidal zone of Dalian

In this study, the biological and chemical diversity of 8 symbiotic marine fungal strains, with strong cytotoxicity against brine shrimp larvae, were investigated by nucleotide sequencing, morphology and cluster analysis of HPTLC fingerprint. These strains were identified by ITS rDNA sequencing, phylogenic analysis, and morphology to be Hypocrea lixii, Chaetomium globosum, Aspergillus fumigatus, Asp. clavatus and Alternaria sp. Their differences in secondary metabolites were shown by cluster analysis of digitalized colors of HPTLC spots, a newly developed method, which produced a similar dendrogram with that of ITS cluster analysis. Furthermore, this method can fully display intraspecific differences and even the remarkable difference in Aspergillus strains which goes beyond the boundary between genera. Their biological-chemical diversity may be the basis of their potent cytotoxicity and implies their potential in producing diversified antitumor or pesticidal constituents.

Zoosporicidal metabolites from an endophytic fungus Cryptosporiopsis sp. of Zanthoxylum leprieurii

Two polyketides, cryptosporiopsin A (1) and hydroxypropan-2',3'-diol orsellinate (3), and a natural cyclic pentapeptide (4), together with two known compounds were isolated from the culture of Cryptosporiopsis sp., an endophytic fungus from leaves and branches of Zanthoxylum leprieurii (Rutaceae). The structures of these metabolites were elucidated on the basis of their spectroscopic and spectrometric data. Cryptosporiopsin A and the other metabolites exhibited motility inhibitory and lytic activities against zoospores of the grapevine downy mildew pathogen Plasmopara viticola at 10-25μg/mL. In addition, the isolated compounds displayed potent inhibitory activity against mycelial growth of two other peronosporomycete phytopathogens, Pythium ultimum, Aphanomyces cochlioides and a basidiomycetous fungus Rhizoctonia solani. Weak cytotoxic activity on brine shrimp larvae was observed.

Cytotoxicity of antimalarial plant extracts from Kenyan biodiversity to the brine shrimp, Artemia salina L. (Artemiidae)

&lt;em&gt;Artemia salina&lt;/em&gt; (Artemiidae), the brine shrimp larva, is an invertebrate used in the alternative test to determine toxicity of chemicals and natural products. In this study the medium lethal concentration fifty (LC50 values) of 45 antimalarial plant extracts and positive controls, cyclophosphamide and etoposide were determined using &lt;em&gt;Artemia salina&lt;/em&gt; (Artemiidae). Out of the 45 organic extracts screened for activity against &lt;em&gt;Artemia salina &lt;/em&gt;larvae, 23 (51%) of the crude extracts demonstrated activity at or below 100 μg/mL, and were categorized as having strong cytotoxic activity, 18 (40%) of the crude extracts had LC50 values between 100 μg/mL and 500 μg/mL, and were categorized as having moderate cytotoxicity, 2 (4.5%) of the crude extracts had LC50 values between 500 μg/mL and 1000 μg/mL, and were considered to have weak cytotoxic activity, while 2 (4.5%) of the crude extracts had LC50 values greater than 1000 μg/mL and were considered to be non toxic. Approximately 20% (9) of the aqueous extracts demonstrated activity at or below 100 g/mL and were considered to have strong cytotoxic activity, 40% (18) of the screened aqueous crude extracts had LC50 values between 100 μg/mL and 500 μg/mL and were considered to be moderately cytotoxic, 16% (7) of the crude extracts had LC50 values between 500 μg/mL and 1000 μg/mL and were considered to have weak cytotoxic activity while 24% (11) of the aqueous extracts had LC50 values greater than 1000μg/mL and were categorized as non toxic The positive controls, cyclophosphamide and etoposide exhibited strong cytotoxicity with LC50 values of 95 μg/mL and 6 μg/mL respectively in a 24 hour lethality study, validating their use as anticancer agents. In the current study, 95.5% of all the screened organic extracts and 76% of the investigated aqueous extracts demonstrated LC50 values &amp;lt;1000 g/mL, indicating that these plants could not make safe anti-malarial treatments. This calls for dose adjustment amongst the community using the plant extracts for the treatment of malaria and chemical investigation for isolation of bioactive compounds responsible for the observed toxicity.

A Quorum Sensing-Disrupting Brominated Thiophenone with a Promising Therapeutic Potential to Treat Luminescent Vibriosis

Vibrio harveyi is amongst the most important bacterial pathogens in aquaculture. Novel methods to control this pathogen are needed since many strains have acquired resistance to antibiotics. We previously showed that quorum sensing-disrupting furanones are able to protect brine shrimp larvae against vibriosis. However, a major problem of these compounds is that they are toxic toward higher organisms and therefore, they are not safe to be used in aquaculture. The synthesis of brominated thiophenones, sulphur analogues of the quorum sensing-disrupting furanones, has recently been reported. In the present study, we report that these compounds block quorum sensing in V. harveyi at concentrations in the low micromolar range. Bioluminescence experiments with V. harveyi quorum sensing mutants and a fluorescence anisotropy assay indicated that the compounds disrupt quorum sensing in this bacterium by decreasing the ability of the quorum sensing master regulator LuxR to bind to its target promoter DNA. In vivo challenge tests with gnotobiotic brine shrimp larvae showed that thiophenone compound TF310, (Z)-4-((5-(bromomethylene)-2-oxo-2,5-dihydrothiophen-3-yl)methoxy)-4-oxobutanoic acid, completely protected the larvae from V. harveyi BB120 when dosed to the culture water at 2.5 µM or more, whereas severe toxicity was only observed at 250 µM. This makes TF310 showing the highest therapeutic index of all quorum sensing-disrupting compounds tested thus far in our brine shrimp model system.

Evaluation of Acute Toxicity of Crude Plant Extracts from Kenyan Biodi-versity using Brine Shrimp, Artemia salina L. (Artemiidae)

Purpose: Artemia salina L. (Artemiidae), the brine shrimp larva, is an invertebrate used in the alternative test to determine toxicity of chemicals and natural products. In this study, we aimed to evaluate the acute toxicity of crude plant extracts using A.salina. Materials and methods: In this study the Medium Lethal Concentrations (LC50 values) of 45 antimalarial plant extracts and positive controls, cyclophosphamide and etoposide were determined using A. salina. Results: Out of the 45 organic extracts screened for activity against A. salina larvae, 23 (51%) of the crude extracts dem- onstrated activity at or below 100 µg/ml, and were categorized as having strong cytotoxic activity, 18 (40%) of the crude extracts had LC50 values between 100 µg/ml and 500 µg/ml, and were categorized as having moderate cytotoxicity, 2 (4.5%) of the crude extracts had LC50 values between 500 µg/ml and 1000 µg/ml, and were considered to have weak cyto- toxic activity, while 2 (4.5%) of the crude extracts had LC50 values greater than 1000 µg/ml and were considered to be non toxic. Approximately 9 (20%) of the aqueous extracts demonstrated activity at or below 100 μg/ml and were consid- ered to have strong cytotoxic activity, 18 (40%) of the screened aqueous crude extracts had LC50 values between 100 µg/ml and 500 µg/ml and were considered to be moderately cytotoxic, 7 (16%) of the crude extracts had LC50 values be- tween 500 µg/ml and 1000 µg/ml and were considered to have weak cytotoxic activity while 11 (24%) of the aqueous ex- tracts had LC50 values greater than 1000 µg/ml and were categorized as non toxic. The positive controls, cyclophos- phamide and etoposide exhibited strong cytotoxicity with LC50 values of 95 µg/ml and 6 µg/ml respectively in a 24 hour lethality study, validating their use as anticancer agents. Conclusions: In the current study, 95.5% of all the screened organic extracts and 76% of the investigated aqueous extracts demonstrated LC50 values <1000 μg/ml, indicating that these plants could not make safe antimalarial treatments. This calls for dose adjustment amongst the community using the plant extracts for the treatment of malaria and chemical investiga- tion for isolation of bioactive compounds responsible for the observed toxicity.

Effects of chronic cocaine in rat C6 astroglial cells

Investigations with astroglial cells carry equal importance as those with neurons in drug abuse studies. The present study was aimed to investigate the effect of chronic cocaine administration on cell viability, nitric oxide (NO) production, general respiratory status of mitochondria and total protein levels in rat astroglioma cells after 24 h of treatment. In addition, the effect of cocaine was assessed for 24 h on brine shrimp larvae in order to study their sensitivity to the drug. It was observed that cocaine caused a significant dose-dependent decrease in astroglial cell viability with an LC50 of 4.717 mM. It was found that cocaine did not induce or inhibit NO production in the cells. Evaluation of mitochondrial dehydrogenase activity in terms of formazan production in astroglial cells indicated that cocaine significantly interfered with the general respiratory status of mitochondria with an ED50 of 6.153 mM. Furthermore, cocaine was shown to deplete the total protein levels in the cells with an ED50 of 5.435 mM. In vivo study with brine shrimp larvae showed that these larvae were highly sensitive to cocaine with an ED50 of 2.41 mM. In summary, our findings suggest that cocaine-induced cytotoxicity in the cells was non-specific. The cumulative effect arising from the significant loss of respiration and total cellular proteins is the cause of astroglial cell death.

Toxic lanostane triterpenes from the basidiomycete Ganoderma amboinense

A highly oxygenated new lanostane triterpene, designated as ganodermacetal (1), together with 15 related known compounds, has been isolated from the fruiting bodies of the basidiomycetous fungus Ganoderma amboinense. The structure of the new metabolite was elucidated on the basis of extensive spectroscopic analyses and chemical transformations. Ganodermacetal (1) possesses an uncommon 7,15-acetonide unit. Some of the isolated compounds showed significant toxic activity against brine shrimp larvae.

Monitoring of Vibrio harveyi quorum sensing activity in real time during infection of brine shrimp larvae

Quorum sensing, bacterial cell-to-cell communication, has been linked to the virulence of pathogenic bacteria. Indeed, in vitro experiments have shown that many bacterial pathogens regulate the expression of virulence genes by this cell-to-cell communication process. Moreover, signal molecules have been detected in samples retrieved from infected hosts and quorum sensing disruption has been reported to result in reduced virulence in different host-pathogen systems. However, data on in vivo quorum sensing activity of pathogens during infection of a host are currently lacking. We previously reported that quorum sensing regulates the virulence of Vibrio harveyi in a standardised model system with gnotobiotic brine shrimp (Artemia franciscana) larvae. Here, we monitored quorum sensing activity in Vibrio harveyi during infection of the shrimp, using bioluminescence as a read-out. We found that wild-type Vibrio harveyi shows a strong increase in quorum sensing activity early during infection. In this respect, the bacteria behave remarkably similar in different larvae, despite the fact that only half of them survive the infection. Interestingly, when expressed per bacterial cell, Vibrio harveyi showed around 200-fold higher maximal quorum sensing-regulated bioluminescence when associated with larvae than in the culture water. Finally, the in vivo quorum sensing activity of mutants defective in the production of one of the three signal molecules is consistent with their virulence, with no detectable in vivo quorum sensing activity in AI-2- and CAI-1-deficient mutants. These results indicate that AI-2 and CAI-1 are the dominant signals during infection of brine shrimp.

Two New Coumarins from Micromelum falcatum with Cytotoxicity and Brine Shrimp Larvae Toxicity

Two new coumarins, 7-methoxy-8-(2-hydroxmethyl-1-O-isovaleryl-4-butenyl)-coumarin (1) and 7-methoxy-8-(1-hydroxy-2-O-β-glucopyranosyl-3-methyl-4-butene-1-yl)coumarin (2), and twelve known coumarins 3–14 were isolated from the stem bark of Micromelum falcatum. The structures of compounds 1–14 were elucidated by extensive spectroscopic data analyses. The toxicity of compounds 1–14 was tested using a brine shrimp assay and in vitro antiproliferative assay against mammary cancer (F10) and lung cancer (HvEvc) cell lines by the MTT method. Some compounds had moderate activities. All compounds were also tested against the microorganisms Bacillus subtilis, Bacillus thuringiensis and Escherichia coli, but no activity was observed.

Paeciloside A, a New Antimicrobial and Cytotoxic Polyketide from Paecilomyces sp. Strain CAFT156

Paeciloside A (1) and eight known compounds, acremoauxin A (2), farinosones A (3) and B (4), 1,5-dideoxy-3-C-methyl-arabitol (5), ergosterol, ergosterol peroxide, cerebroside C, and adenosine, were isolated from cultures of Paecilomyces sp. CAFT156, an endophytic fungus occurring in Enantia chlorantha Oliv (Annonaceae) leaves. The structure of the new compound 1 was elucidated using MS, UV, 1D and 2D NMR experiments, while its absolute configuration was established by subsequent single-crystal X-ray diffraction analysis using copper Kα radiation and invariom nonspherical scattering factors. Paeciloside A (1) and compounds 2, 4, and 5 displayed inhibitory effects on two gram-positive bacteria, Bacillus subtilis and Staphylococcus aureus at a concentration of 40 µg per paper disk and moderate cytotoxicity towards brine shrimp larvae (Artemia salina). This study presents the first report on an endophytic fungus isolated from E. chlorantha Oliv and its natural products.

Cytotoxic Effect of Five Medicinal Plants Extracts Using Brine Shrimp (&lt;i&gt;Artemia salina&lt;/i&gt;) TEST

Medicinal plants like Curcuma longa, Curcuma zedoaria, Streblus asper, Enhydra fluctunas and Scoparia dulcis are commonly available in Bangladesh. This study investigated the effect of ethanol extracts from these five medicinal plants on the viability of Brine Shrimp larvae. The lethal concentrations (LC50 values) of the extracts were determined. LC50 values of C. longa, C. zedoaria and S. asper were less than 250?g/ml and that of E. fluctunas was greater than 250 ?g/ml. Among five plants tested, one (S. dulcis) appear to be inactive. Results indicate that C. longa, C. zedoaria and S. asper could be potential sources of bio-active compounds DOI: http://dx.doi.org/10.3329/bjsir.v47i1.10724 Bangladesh J. Sci. Ind. Res. 47(1), 133-136, 2012

Phytochemical composition, cytotoxicity and in vitro antiplasmodial activity of fractions from Alafia barteri olive (Hook F. Icon)-Apocynaceae

The ethanolic extract of Alafia barteri (Hook F. Icon) was dissolved in distilled water and successively partitioned in n-hexane, chloroform, ethyl acetate and n-butanol. The fractions were evaluated for phytochemical composition, lethality against brine shrimp larvae and in vitro antiplasmodial activity against Plasmodium falciparum strain. The obtained results revealed that the roots and leaf extracts of A. barteri exhibited broad spectrum of antiplasmodial activity (IC50 1.5±0.7–6.2±0.80μg/mL). The aqueous leaf fractions displayed the most potent antiplasmodial activity with an IC50 value of 1.5±0.7μg/mL, which is comparable to reference antimalarial drug (IC50 value of 1.3±0.2μg/mL). The leaf fractions displayed higher activity than the root extracts. The highest minimum lethal concentration (105.2±0.8ppm) was exhibited by the aqueous leaf extract followed closely by the root extract (120.2±1.1ppm). The leaf extracts contained higher polyphenols (45.3±0.85mgGAE/g) and flavonoids (18.10±0.2mgCTE/g) than the root extracts. The n-hexane and EtOAc extracts/fractions displayed lower activity on brine shrimp larvae.

Biologically active abietane and ent-kaurane diterpenoids and other constituents from Erythroxylum suberosum

Bioassay-guided fractionation of the ethanol extract from the branches of Erythroxylum suberosum, which was toxic to brine shrimp larvae, afforded five diterpenes bearing abietane and ent-kaurane-type skeletons from an active fraction. From these, four were new, 7-oxo-16-hydroxy-abiet-15(17)-en-19-al, 16-hydroxyabiet-15(17)-en-7-one, 7α,16-dihydroxy-abiet-15(17)-en-19-al and ent-12α-hydroxy-kaur-16-en-19-al, while methyl ent-7α,15β-dihydroxy-kaur-16-en-19-oate is reported for the first time as a natural product. This is also the first reported occurrence of abietane-type diterpenes in the genus Erythroxylum. The flavonoid ombuin-3-rutinoside was isolated from an inactive fraction, while rutin (quercetin-3-rutinoside) was obtained from the non-toxic ethanol extract of the leaves. The structures of the new and known compounds were established by analyses of 1D- and 2D-NMR and mass spectrometry data.

Evaluation of antifungal activity from Bacillus strains against Rhizoctonia solani

In this study, 30 bacterial strains isolated from marine biofilms were screened for their antifungal activity against Rhizoctonia solani by dual culture assay. Two bacterial strains, Bacillus subtilis and Bacillus cereus , showed a clear antagonism against R. solani on potato dextrose agar (PDA) medium. The antagonistic activity of B. subtilis against this pathogen persisted after one month of co-culture. An in vitro antagonistic assay using potato tuber slices was set up allowing both the screening of bacteria for their biocontrol proprieties and for their rotting effect on plant tissues. Potato tuber slices treated with B. cereus showed a heavy tissues rotting, probably due to the secretion of several enzymes. Whereas, B. subtilis did not show any tissues rotting on treated potato tuber slices. In addition, the potato tuber slices treated with B. subtilis and subsequently inoculated with R. solani agar culture showed a reduced fungal infection in comparison to the control tuber slices inoculated with the pathogen only. The crude extract of B. subtilis strain culture in Luria-Bertani (LB) medium at 48 h of incubation showed a high antifungal activity against R. solani growth and no cytotoxic effect on Brine shrimp larvae. Key words: Antagonism, Bacillus spp., biological control, black scurf of potato, marine biofilms.

A method for the specific detection of resident bacteria in brine shrimp larvae

In this study, we describe an easy but efficient method to specifically target the intestinal resident microbiota in brine shrimp larvae during DGGE analysis, hereby excluding the interference of both transient (luminal) bacteria and body surface bacteria. This effective technique has several advantages over alternative methods, with respect of ease of use and rapidity.

A New Cytotoxic Sesquiterpene Quinone Produced by Penicillium sp. F00120 Isolated from a Deep Sea Sediment Sample

A new fungal strain, displaying strong toxic activity against brine shrimp larvae, was isolated from a deep sea sediment sample collected at a depth of 1300 m. The strain, designated as F00120, was identified as a member of the genus Penicillium on the basis of morphology and ITS sequence analysis. One new sesquiterpene quinone, named penicilliumin A (1), along with two known compounds ergosterol (2) and ergosterol peroxide (3), were isolated and purified from the cultures of F00120 by silica gel column, Sephadex LH-20 column, and preparative thin layer chromatography. Their structures were elucidated by detailed nuclear magnetic resonance (NMR) and mass spectroscopic (MS) analysis as well as comparison with literature data. The new compound penicilliumin A inhibited in vitro proliferation of mouse melanoma (B16), human melanoma (A375), and human cervical carcinoma (Hela) cell lines moderately.

Anti-Mycobacterial and Toxicity Activities of Some Priority Medicinal Plants from Lake Victoria Basin, Tanzania

Aims: This study has evaluated ethanol extracts from five medicinal plants selected through ethnobotanical study from Lake Victoria basin, Tanzania for their in vitro antimycobacterial activity against two Mycobacterium species and cytotoxicity against brine shrimp larvae. Study Design: Laboratory experimental tests. Place and Duration of Study: Institute of Traditional Medicine, Muhimbili University of Health and Allied Sciences, P.O. Box 65001, Dar es Salaam, Tanzania, between July 2010 and July 2011. Methodology: Five medicinal plants were selected from the priority list obtained from Lake Victoria basin, Tanzanian side. Collection, processing and drying of plant samples were done in the field with the assistance of a botanist while extraction and concentration of plant samples to obtain crude extracts were done in the laboratory following standard procedures. The plants included in this study are Antidesma membranaceum, Crassocephalum manii, Entada abyssinica, Croton dichogamus and Rubia cordifolia. The two fold microdilution method was used to determine the MIC values of extracts against Research Article European Journal of Medicinal Plants, 2(2): 125-131, 2012 126 two Mycobacterium marker strains (Mycobacterium indicus pranii and Mycobacterium madagascariense). The cytotoxicity of plant extract was evaluated against brine shrimp larvae. Furthermore, the extracts were screened phytochemicaly to establish the group of compounds responsible for the activity. Results: Among the tested extracts, the stem bark of A. membranaceum and C. manii showed moderate to mild activity against M. indicus pranii (MIC = 0.3125 mg/ml) and M. madagascariense (MIC = 0.625 mg/ml) respectively. Furthermore, A. membranaceum exhibited significant toxicity activity with LC50 value of 36.134 μg/ml against brine shrimp larvae. Other plants were moderately active when tested in vitro against the above organisms. Phytochemical screening of extracts indicated the presence of different classes of compounds. Conclusion: This study has shown the potential of the priority medicinal plant extracts to be the source of possible lead compounds and anti‐TB drug candidates needed for the management of Tuberculosis. Isolation of active principles from active fractions will be further undertaken.