scholarly journals Enzymatic activity and brine shrimp lethality of venom from the large brown spitting cobra (Naja ashei) and its neutralization by antivenom

2020 ◽  
Author(s):  
Mitchel Otieno Okumu ◽  
James Mucunu Mbaria ◽  
Joseph Kangangi Gikunju ◽  
Paul Gichohi Mbuthia ◽  
Vincent Odongo Madadi ◽  
...  

Abstract Objective: Naja ashei is a snake of medical importance in Kenya, Ethiopia, Somalia, Uganda, and Tanzania. Little is known about the enzymatic (snake venom phospholipases A2; svPLA2’s) and toxic (lethal) activities of Naja ashei venom and crucially, the safety and capacity of available antivenom to neutralize these effects. This study aimed to determine the enzymatic and toxic activities of Naja ashei venom and the capacity of Indian and Mexican manufactured antivenoms to neutralize these effects. The protein content of the venom and the test antivenoms were also evaluated. A 12-point log concentration-response curve (0.5-22.5 µg/mL) was generated on an agarose-egg yolk model to predict the svPLA2 activity of the venom. The toxicity profiles of the venom and antivenoms were evaluated in the brine shrimp lethality assay. Lowry’s method was used for protein estimation. Results: Low and intermediate concentrations of the venom exhibited similar svPLA2 activities. The same was true for concentrations >15 µg/mL. Intermediate and high doses of the venom exhibited similar mortalities in brine shrimp and test antivenoms were generally non-toxic but poorly neutralized svPLA2 activity. Mexican manufactured antivenom had lower protein content but neutralized venom-induced brine shrimp lethality much more effectively than Indian manufactured antivenom.

2020 ◽  
Author(s):  
Mitchel Otieno Okumu ◽  
James Mucunu Mbaria ◽  
Joseph Kangangi Gikunju ◽  
Paul Gichohi Mbuthia ◽  
Vincent Odongo Madadi ◽  
...  

Abstract Objective: Naja ashei is a snake of medical importance in Kenya, Ethiopia, Somalia, Uganda, and Tanzania. Little is known about the enzymatic (snake venom phospholipases A2; svPLA2’s) and toxic (lethal) activities of Naja ashei venom and crucially, the safety and capacity of available antivenom to neutralize these effects. This study aimed to determine the enzymatic and toxic activities of Naja ashei venom and the capacity of Indian and Mexican manufactured antivenoms to neutralize these effects. The protein content of the venom and the test antivenoms were also evaluated. A 12-point log concentration-response curve (0.5-22.5 µg/mL) was generated on an agarose-egg yolk model to predict the svPLA2 activity of the venom. The toxicity profile of the venom and antivenoms was evaluated in the brine shrimp lethality assay. Lowry’s method was used for protein estimation. Results: Low and intermediate concentrations of the venom exhibited similar svPLA2 activities. The same was true for concentrations >15µg/mL. Intermediate and high doses of the venom exhibited similar mortalities in brine shrimp and test antivenoms were generally non-toxic but poorly neutralized svPLA2 activity. Mexican manufactured antivenom had lower protein content but neutralized venom-induced brine shrimp lethality much more effectively than Indian manufactured antivenom.


2009 ◽  
Vol 2 (1) ◽  
pp. 169-177
Author(s):  
M. A. Rahman ◽  
M. S. Rana ◽  
M. M. Zaman ◽  
S. A. Uddin ◽  
R. Akter

The antioxidant, antibacterial and cytotoxic properties of methanol extract of Urtica crenulata (syn: Laportea crenulata Gaud) stem has been investigated in the present study. The antioxidant and cytotoxic activity of the Urtica crenulata methanol extract were assessed by DPPH free radical scavenging method and brine shrimp lethality bioassay method, respectively. The antibacterial activity of the plant extract (500 μg/disc) was also carried out by disc diffusion technique. Stem extract showed DPPH (2,2-diphenyl-1-picrylhydrazyl) free radical scavenging effect compared with ascorbic acid. IC50 value of ascorbic acid and stem extract was found 14.72 μg/ml and 1468.9 μg/ml, respectively. In antibacterial experiment, Urtica crenulata stem extract showed 8, 14 and 10 mm of diameter of zone inhibition against Salmonella typhi, Shigella flexneri and Shigella sonnei, respectively and 9 and 8 mm of diameter of zone inhibition against Bacillus subtilis and Bacillus cereus but no activity was observed against Staphylococcus aureus. In brine shrimp lethality assay, the LC50 value of the extract was found 104.0 μg/ml, which indicates that the extract has high cytoxic effect. The present study demonstrates that methanol extract of Urtica crenulata stem has significant cytotoxic effect. The extract also showed some moderate antibacterial and minimum significant antioxidant effects.  Keywords: Urtica crenulata;  Antioxidant; Antibacterial; Cytotoxic; BHT. © 2010 JSR Publications. ISSN: 2070-0237 (Print); 2070-0245 (Online). All rights reserved. DOI: 10.3329/jsr.v2i1.2872             J. Sci. Res. 2 (1), 169-177 (2010) 


Proceedings ◽  
2019 ◽  
Vol 41 (1) ◽  
pp. 47
Author(s):  
Sandeep Waghulde ◽  
Mohan K. Kale ◽  
VijayR. Patil

The present study was conducted to test for in vivo Brine Shrimp Lethality Assay (BSLA) of the Aqueous and ethanolic extracts of Annona reticulata with Allium fistolisum and Brassica oleraceaeand correlate cytotoxicity results with known pharmacological activities of the plants. Cytotoxicity was evaluated in terms of LC50 (lethality concentration). Ten nauplii were added into three replicates of each concentration of the plant extract. After 24 h the surviving brine shrimp larvae were counted and LC50 was assessed. Results showed that the extracts of Annona reticulata with Allium fistolisumand Brassica oleraceaewere potent against the brine shrimp when compared alone with combined extracts. It indicated that bioactive components are present in these plants that could be accounted for its pharmacological effects. Thus, the results support the uses of these plant species in traditional medicine.


2019 ◽  
Vol 2019 ◽  
pp. 1-8 ◽  
Author(s):  
Jahangir Khan Achakzai ◽  
Muhammad Anwar Panezai ◽  
Muhammad Ayub Kakar ◽  
Abdul Manan Kakar ◽  
Shahabuddin Kakar ◽  
...  

In this study, GC-MS analysis has shown that whole plant butanol fraction of rheum ribes (WBFRR) comprises of 21 compounds which exhibited anticancer (MCF-7) activity having IC50 value of 36.01± 0.26. MTT assay (MCF-7), Oxidative Burst assay using chemiluminescence technique, and B-Hatching techniques were the methods used for anticancer MCF-7, anti-inflammatory, and Brine Shrimp Lethality Assay (BSLA). GC-MS was used for structural elucidation. Whole plant methanol extract of rheum ribes (WMERR), whole plant n-hexane fraction of rheum ribes (WHFRR), and whole plant aqueous fraction of rheum ribes (WAFRR) were inactive against anticancer (MCF-7) cell line. Whole plant methanol extract of rheum ribes (WMERR), whole plant aqueous fraction of rheum ribes (WAFRR) and whole plant butanol fraction of rheum ribes (WBFRR) showed anti-inflammatory activity on ROS having IC50 value of 23.2±1.9, 24.2±2.7 and 12.0±0.6. Whole plant butanol fraction of rheum ribes (WBFRR) showed Brine Shrimp Lethality with LD50 693.302 while whole plant methanol extract of rheum ribes (WMERR) and whole plant aqueous fraction of rheum ribes (WAFRR) showed high lethality at highest concentration. This study revealed that whole plant butanol fraction of rheum ribes (WBFRR) exhibited significant anticancer (MCF-7) activity. In the near future, the constituent of whole plant butanol fraction of rheum ribes (WBFRR) can be the alternative drug against MCF-7 cell line with least toxicity and side effects.


Author(s):  
Anik Barua ◽  
Md. Rabiul Hossain ◽  
Labonno Barua ◽  
Abu Sadat Md. Sayem ◽  
Md. Zainul Abedin ◽  
...  

Aims: Tinospora cordifolia is a herb used in the traditional folk medicine due to their health benefits. Studies reported the biochemical composition and some biological properties of the plant stem. The aim of the current study was to investigate the antimicrobial and cytotoxic properties from the extract of Tinospora cordifolia stem. Place and Duration of Study: The study was accomplished by eleven months in the Department of Pharmacy, University of Science and Technology Chittagong (USTC), Bangladesh.  Methodology: Presence of phytochemical constituents such as alkaloids, flavonoids and tannins, saponins were evaluated. Antimicrobial activity of the stem extract was assayed separately using an agar diffusion method against some gram negative and gram positive bacteria. The cytotoxic potential of the stem extract was evaluated by the Brine-Shrimp Lethality Assay mathod.  Results: The zone of inhibition formed by the methanolic extract in the antimicrobial assays was significant. In addition, cytotoxicity of the extract was found to be significant as compared to the standard gallic acid in Brine-Shrimp lethality assay. Conclusion: The methanolic extract of Tinospora cordifolia may possess significant antimicrobial property which requires more studies to isolate the specific bioactive compound for developing a new antimicrobial agent from this plant.


2014 ◽  
Vol 60 (01) ◽  
pp. 9-18 ◽  
Author(s):  
Mentor R. Hamidi ◽  
Blagica Jovanova ◽  
Tatjana Kadifkova Panovska

Many natural products could serve as the starting point in the development of modern medicines because of their numerous biological and pharmacological activities. However, some of them are known to carry toxicological properties as well. In order to achieve a safe treatment with plant products, numerous research studies have recently been focused on both pharmacology and toxicity of medicinal plants. Moreover, these studies employed efforts for alternative biological assays. Brine Shrimp Lethality Assay is the most convenient system for monitoring biological activities of various plant species. This method is very useful for preliminary assessment of toxicity of the plant extracts. Rapidness, simplicity and low requirements are several advantages of this assay. However, several conditions need to be completed, especially in the means of standardized experimental conditions (temperature, pH of the medium, salinity, aeration and light). The toxicity of herbal extracts using this assay has been determined in a concentration range of 10, 100 and 1000 µg/ml of the examined herbal extract. Most toxicity studies which use the Brine Shrimp Lethality Assay determine the toxicity after 24 hours of exposure to the tested sample. The median lethal concentration (LC50) of the test samples is obtained by a plot of percentage of the dead shrimps against the logarithm of the sample concentration. LC50 values are estimated using a probit regression analysis and compared with either Meyer’s or Clarkson’s toxicity criteria. Furthermore, the positive correlation between Meyer’s toxicity scale for Artemia salina and Gosselin, Smith and Hodge’s toxicity scale for higher animal models confirmed that the Brine Shrimp Lethality Assay is an excellent predictive tool for the toxic potential of plant extracts in humans.


2002 ◽  
Vol 16 (2) ◽  
pp. 197-198 ◽  
Author(s):  
Prashanth D'Souza ◽  
Mundkinajeddu Deepak ◽  
Padmaja Rani ◽  
Sandhya Kadamboor ◽  
Anjana Mathew ◽  
...  

PeerJ ◽  
2018 ◽  
Vol 6 ◽  
pp. e4804 ◽  
Author(s):  
Kumudu R.V. Bandara ◽  
Chayanika Padumadasa ◽  
Dinithi C. Peiris

Passiflora suberosa L. belonging to the family Passifloraceae is an important medicinal plant used in traditional medicinal system in Sri Lanka to treat diabetes, hypertension and skin diseases. We extracted P. suberosa leaves under reflux conditions using different solvents (hexane, chloroform, methanol and water), then subjected to phytochemical screening. Alkaloids, flavonoids and saponins and saponins and anthraquinones were present in hexane and chloroform extracts. Alkaloids, unsaturated sterols, triterpenes, saponins, flavonoids and tannins were observed in both methanol and aqueous extracts. Proanthocyanidins were observed only in the aqueous extract. Hence, aqueous and methanol extracts with most classes of phytochemicals present were subjected to antimicrobial, antioxidant, antihaemolytic activities and Brine shrimp lethality studies. Antibacterial activity and minimum inhibition concentrations were evaluated using three Gram-positive (Bacillus subtilis, Staphylococcus aureus and Enterococcus faecium) and three Gram-negative bacteria (Pseudumonas aeruginosa, Salmonella typhimuriam and Escherichia coli). The results indicated that only the methanol extract of P. suberosa exhibited antibacterial activities against all the strains of Gram-negative and Gram-positive bacterial with stronger activity against Gram-negative bacteria. DPHH (2,2-diphenyl-1-picrylhydrazy) scavenging assay was adopted to evaluate antioxidant properties while antihaemolytic and toxic activities were studied respectively using cow blood and Brine shrimp lethality assay. The IC50 values of the aqueous extract in both antioxidant and antihaemolytic assays were significantly lower than the standard ascorbic acid. Similar results were observed in the Brine shrimp lethality assay. In conclusion both aqueous and methanol extracts of P. suberosa leaves showed the presence of majority of phytochemicals including proanthocyanidins. Antibacterial activity was obtained only for methanol extract with better activity against Gram-negative bacteria. The aqueous extract showed better antioxidant, antihaemolytic and toxic activities than the methanol extract and their respective standards. Further investigations on the chemical composition and possible isolation of active ingredients is warranted.


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