Therapeutic Values of Earthworm Species Extract from Azad Kashmir as Anticoagulant, Antibacterial, and Antioxidant Agents

Abstract

Aims. Current research aimed to explore the therapeutic values of different earthworms as antibacterial, anticoagulant, and antioxidant agents. Methods. Ten different earthworms, i.e., Amynthas corticis, Amynthas gracilis, Pheretima posthuma, Eisenia fetida, Aporrectodea rosea, Allolobophora chlorotica, Aporrectodea trapezoides, Polypheretima elongata, Aporrectodea caliginosa, and Pheretima hawayana, were collected and screened for biological activities. Antibacterial effect analysis of earthworm species was done against fourteen bacterial pathogens, i.e., Escherichia coli, Serratia marcescens, Streptococcus pyogenes, Staphylococcus epidermidis, Staphylococcus aureus, Klebsiella pneumoniae, Pseudomonas aeruginosa (1), Salmonella typhimurium, Shigella flexneri, Enterobacter amnigenus, Serratia odorifera, Pseudomonas aeruginosa (2), Staphylococcus warneri, and Lactobacillus curvatus, via agar well diffusion, crystal violet, MTT, agar disc diffusion, and direct bioautography assays. Antioxidant potential was evaluated through ABTS and DPPH assays. Lipolytic, proteolytic, and amylolytic assays were done for lipase, protease, and amylase enzymes confirmation. In vitro anticoagulant effects were examined in the blood samples by measuring prothrombin time. Results. Results revealed that all earthworm extracts showed the inhibition of all tested bacterial pathogens except P. aeruginosa (1), P. aeruginosa (2), S. warneri, and L. curvatus. The maximum zone of inhibition of E. coli was recorded as 14.66 ± 0.57 mm by A. corticis, 25.0 ± 0.0 mm by P. posthuma, 20.0 ± 0.0 mm by E. fetida, and 20.0 ± 0.0 mm by A. trapezoid. Cell proliferation, biofilm inhibition, the synergistic effect of extracts along with antibiotics, and direct bioautography supported the results of agar well diffusion assay. Similarly, P. hawayana, A. corticis, A. caliginosa, and A. trapezoids increase the prothrombin time more efficiently compared to other earthworms. A. corticis, A. gracilis, A. rosea, A. chlorotica, P. elongata, and A. trapezoides showed maximum DPPH scavenging potential effect. Conclusions. The coelomic fluid of earthworms possessed several bioactive compounds/enzymes/antioxidants that play an important role in the bacterial inhibition and act as anticoagulant agents. Therefore, the development of new therapeutic drugs from invertebrates could be effective and potential for the prevention of the emergence of multidrug-resistant bacteria.

Antibiofilm Potential of Coelomic Fluid and Paste of Earthworm Pheretima posthuma (Clitellata, Megascolecidae) against Pathogenic Bacteria

“…At the end, it was observed that the results established a directly proportional relationship between PBP and PCF concentrations and the biofilm inhibition percentage. Similarly, Mustafa et al [ 44 ] used the earthworms’ extract to check its effect on biofilm inhibition, cell proliferation, and the synergistic effect. They found that the earthworm extract inhibited the biofilm synthesis compared to the control.…”

“…Moreover, the extract of A. gracilis also reduced the biofilm synthesis for S. aureus and S. pyogenes , while the extract of P. posthuma inhibited the biofilm formation for E. coli and K. pneumoniae. Likewise, body extracts of E. fetida reduced the biofilm development for P. aeruginosa [ 44 ]. Biofilm formation is based on various factors and their correlations, such as adhesions, growth, extracellular-binding proteins in matrix, biofilm architecture, and cell to cell communication, which play a vital role in formation as well as maturation [ 33 ].…”

“…Similarly, Hussain et al [ 17 ] described that earthworm body paste, extract, and coelomic fluid all possess various kinds of bioactive agents, including proteases (proteins or peptides), metabolites, metal-binding proteins, active proteins (such as lysenin, lysozyme, and eiseniapore, etc. ), antimicrobial peptides, coelomic cytolytic factor (CCF and CCF-I), lysenin, fetidin, lumbricin complex, organic acids (i.e., fatty acid), and some organic compounds (e.g., vitamin D and purine and vitamin D) [ 7 , 44 ]. These bioactive compounds might have played a significant role as antibiofilm agents in the current study.…”

Allolobophora caliginosa coelomic fluid and extract alleviate glucocorticoid-induced osteoporosis in mice by suppressing oxidative stress and regulating osteoblastic/osteoclastic-related markers

“…Thus, it has been consumed in various ways; in previous years, earthworm was advised to be edible for anemia and malaria 13 . Recent research dedicated that Allolobophora calignosa, which is an earthworm species that belong to the phylum Annelida 14 , possesses many activities such as antioxidant 11 , anticoagulant, antibacterial and anticancer 15 .…”

Histopathological examination of Allolobophora calignosa efficacy against glucocorticoid-induced osteoporosis in mice

“…The earthworm has proven a competitive medicinal importance in treating various diseases due to its biological activities. The body cavity of earthworms comprises a coelomic fluid, which has anti-inflammatory capabilities (16). Extract of earthworm (Pheretima aspergillum) increases osteoblast activity and inhibits the generation of osteoclasts (17).…”