Abstract
Trichinella spiralis (T. spiralis) is a prevalent foodborne intestinal parasite in many developing countries. Albendazole (ABZ) is the drug of choice for treating trichinosis despite its several drawbacks as its week effect against encapsulated larvae, low bioavailability, and emerging drug resistance. As a result, new anthelmintic agents are required. This study aims to investigate the in vivo and in vitro effects of Punica granatum peels extract (PGPE) on intestinal and muscle phases of T. spiralis. The adult worms and larvae were isolated and cultured with different concentrations of PGPE ranging from 6.75 to 100 µg/ml and measuring the survival rate was done after 1, 3, 18, 24 and 48 h of incubation, followed by scanning electron microscopic (SEM) examination of isolated parasites. For the in vivo experiment, the infected animals were divided into two main groups: intestinal phase group and muscular phase group, each group was subdivided into; infected not treated, infected treated with PGPE, ABZ and combined PGPE and ABZ (6 mice in each). The drug effect was assessed by adults and larvae load. A significant increase in the percentage of dead adult parasite and muscle larvae cultured with PGPE with severe destruction and deformity of the tegument were observed with SEM. Also, a significant reduction of adult parasite number in the intestine and muscle larva number in the diaphragm of infected treated mice in comparison to the control group. This study proved that PGPE has a potential activity against trichinosis, particularly when combined with ABZ, and this could serve as a new agent in trichinosis therapy.
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References
Abdel Aziz AR, AbouLaila MR, Aziz M, Omar MA, Sultan K (2018) In vitro and in vivo anthelmintic activity of pumpkin seeds and pomegranate peels extracts against Ascaridia galli. Beni-Suef Univ J Basic Appl Sci 7(2):231–234. https://doi.org/10.1016/j.bjbas.2018.02.003
Abdel Moneim AE (2012) Evaluating the potential role of pomegranate peel in aluminum-induced oxidative stress and histopathological alterations in brain of female rats. Biol Trace Elem Res 150:328–336
Abdel-Ghaffar F, Semmler M, Al-Rasheid KA, Strassen B, Fischer K, Aksu G, Klimpel S, Mehlhorn H (2011) The effects of different plant extracts on intestinal cestodes and on trematodes. Parasitol Res 108:979–984
Abdel-Hafeez EH, Ahmed AK, Abdellatif MZM, Kamal AM, Toni NDM (2016) The efficacy of pomegranate (Punica granatum) peel extract on experimentally infected rats with Blastocystis Spp. J Infect Dis Preve Med 4:131
Aboelsoued D, Abo-Aziza FAM, Mahmoud MH, Abdel Megeed KN, Abu El Ezz NMT, Abu-Salem FM (2019) Anticryptosporidial effect of pomegranate peels water extract in experimentally infected mice with special reference to some biochemical parameters and antioxidant activity. J Parasit Dis 43:215–228
Ahmed AH, Ejo M, Feyera T, Regassa D, Mummed B, Huluka SA (2020) In vitro anthelmintic activity of crude extracts of Artemisia herba-alba and Punica granatum against Haemonchus contortus. J Parasitol Res 2020:4950196. https://doi.org/10.1155/2020/4950196
Al-Mathal EA, Alsalem MA (2012) Pomegranate (Punica granatum) peel is effective in a murine model of experimental Cryptosporidium parvum. Exp Parasitol 131:350–357
Al-Megrin WA (2016) Efficacy of pomegranate (Punica granatum) peel extract against Hymenolepis nana in infections mice. Biosci Biotech Res Asia 13(1):103–108
Al-Megrin WA (2017) In vivo study of pomegranate (Punica granatum) peel extract efficacy against Giardia lamblia in infected experimental mice. Asian Pac J Trop Biomed 7(1):59–63. https://doi.org/10.1016/j.apjtb.2016.08.018
Anand U, Jacobo-Herrera N, Altemimi A, Lakhssassi NA (2019) comprehensive review on medicinal plants as antimicrobial therapeutics: potential avenues of biocompatible drug discovery. Metabolites 9(11):258. https://doi.org/10.3390/metabo9110258.PMid:31683833
Arun KB, Jayamurthy P, Anusha CV, Mahesh SK, Nisha P (2017) Studies on activity guided fractionation of pomegranate peel extracts and its effect on antidiabetic and cardiovascular protection properties. J Food Process Preserv 41:e13108
Attia RA, Mahmoud AE, Farrag HM, Makboul R, Mohamed ME, Ibraheim Z (2015) Effect of myrrh and thyme on Trichinella spiralis enteral and parenteral phases with inducible nitric oxide expression in mice. Memorias Do Institue Oswaldo Cruz 110(8):1035–1041
Barathikannan K, Venkatadri B, Khusro A, Al-Dhabi NA, Agastian P, Arasu MV et al (2016) Chemical analysis of Punica granatum fruit peel and its in vitro and in vivo biological properties. BMC Complement Altern Med 16(1):264. https://doi.org/10.1186/s12906-016-1237-3.PMid:27476116
Bassiri-Jahromi S, Pourshafie MR, Ardakani EM, Ehsani AH, Doostkam A, Katirae F, Mostafavi E (2018) In vivo comparative evaluation of pomegranate (Punica granatum) peel extract as alternative agents for nystatin against oral candidiasis. Iran J Med Sci 43(3):296–304
Basyoni MM, El-Sabaa AAA (2013) Therapeutic potential of myrrh and ivermectin against experimental Trichinella spiralis infection in mice. Korean J Parasitol 51(3):297
Boonmasawai S, Sungpradit S, Jirapattharasate C, Nakthong C, Piasai L (2017) Effects of alcoholic extract from pomegranate (Punica granatum L.) peels on gastrointestinal nematode egg counts in doe. J App Anim Sci 6(2):27–37
Botura MB, dos Santos JD, da Silva GD, de Lima HG, de Oliveira JV, de Almeida MAO, Batatinha MJM, Branco A (2013) In vitro ovicidal and larvicidal activity of Agave sisalana Perr. (sisal) on gastrointestinal nematodes of goats. Vet Parasitol 192(1–3):211–217. https://doi.org/10.1016/j.vetpar.2012.10.012
Castagna F, Britti D, Oliverio M, Bosco A, Bonacci S, Iriti G et al (2020) In vitro anthelminthic efficacy of aqueous pomegranate (Punica granatum L.) extracts against gastrointestinal nematodes of sheep. Pathogens 9(12):1063. https://doi.org/10.3390/pathogens9121063.PMid:33353177
Dilks CM, Hahnel SR, Sheng Q, Long L, McGrath PT, Andersen EC (2020) Quantitative benzimidazole resistance and fitness effects of parasitic nematode beta-tubulin alleles. Int J Parasitol Drugs Drug Resist 14:28–36. https://doi.org/10.1016/j.ijpddr.2020.08.003.PMid:32858477
Dkhil MA (2013) Anti-coccidial anthelmintic and antioxidant activities of pomegranate (Punica granatum) peel extract. Parasitol Res 112(7):2639–2646. https://doi.org/10.1007/s00436-013-3430-3.PMid:23609599
Dos Santos RL, Dellacqua LO, Delgado NT, Rouver WN, Podratz PL, Lima LC, Piccin MP, Meyrelles SS, Mauad H, Graceli JB, Moyses MR (2016) Pomegranate peel extract attenuates oxidative stress by decreasing coronary angiotensin-converting enzyme (ACE) activity in hypertensive female rats. J Toxicol Environ Health A 79:998–1007
Dunn IJ, Wright KA (1985) Cell injury caused by Trichinella spiralis in the mucosal epithelium of B10A mice. J Parasitol 71(6):757–766
Esmat M, Abdel-Aal AA, Shalaby MA, Fahmy MA, Badawi MA, Elmallawany MA, Magdy M, Afife AA, Abdel Shafi IR (2021) Punica granatum and amygdalin extracts plus cobalamin combined with albendazole reduce larval burden and myositis in experimental trichinosis. Braz J Vet Parasitol 30(4):e012021. https://doi.org/10.1590/S1984-29612021084
Fahmy Z, El-Shennawy A, El-Komy W, Ali E, Abd E-H (2009) Potential antiparasitic activity of pomegranate extracts against schistosomules and mature worms of Schistosoma mansoni: in vitro and in vivo study. Aust J Basic Appl Sci 3(4):4634–4643. https://doi.org/10.1016/s1201-9712(10)60263-9
Glauert AM (1974) Fixation, dehydration and embedding of biological specimens. In: Glauert AM (ed) Practical methods in electron microscopy. Amsterdam, Oxford
Hafez EN, El Kholy WA, Amin MM (2020) The potential protective role of gamma-irradiated vaccine versus Punica granatum treatment against murine trichinellosis. J Radiat Res 13(1):560–567. https://doi.org/10.1080/16878507.2020.1777659
Hettiarachchi G, Samanta SK, Falcinelli S, Zhang B, Moncelet D, Isaacs L et al (2016) Acyclic cucurbit[n]uril-type molecular container enables systemic delivery of effective doses of albendazole for treatment of SK-OV-3 xenograft tumors. Mol Pharm 13(3):809–818. https://doi.org/10.1021/acs.molpharmaceut.5b00723.PMid:26756920
Issa RM, El-Arousy MH, Abd EI-Aal AA (1998) Albendazole: a study of its effect on experimental Trichinella spiralis infection in rats. Egypt J Med Sci 19:281–290
Jabeen N, Anwar S, Mahmood Q, Zia MA, Murtaza G (2015) In vitro anthelmintic efficacy of native plants against Haemonchus contortus. Acta Poloniae Pharmaceutica Drug Res 72(5):1051–1055
Jurenka JS (2008) Therapeutic applications of pomegranate (Punica granatum L.): a review. Altern Med Rev 13(2):128–144
Labsi M, Khelifi L, Mezioug D, Soufli I, Touil-Boukoffa C (2016) Antihydatic and immunomodulatory effects of Punica granatum peel aqueous extract in a murine model of echinococcosis. Asian Pac J of Trop Med 9(3):211–220
Muñoz-Carrillo JL, Muñoz-Escobedo JJ, Maldonado-Tapia CH, Chávez-Ruvalcaba F, Moreno-García MA (2017) Resiniferatoxin lowers TNF-α, NO and PGE2 in the intestinal phase and the parasite burden in the muscular phase of Trichinella spiralis infection. Parasite Immunol 39(1):e12393
Oshiba SF, Yaseein RI, El-Shennawy AM, Aiad HA, El-Wakil EA (2018) In vivo effect of pomegranate (Punica granatum) extracts versus Nitazoxanide drug on the ileum of experimentally infected mice with Cryptosporidium parvum oocysts. J Am Sci 14(2):27–39
Osman GY, Mohamed AH, Salem TA, Elmalawany AM (2013) Immunoparasitological effect of punica granatum in Schistosoma mansoni infected mice. In: 10th International conference on future horizon of environmental sustainable development in arab countries and facing the challenges Sharm El-Sheikh, Egypt, pp 21–24/12/2013
Ozkoc S, Tuncay S, Delibas SB, Akisu C (2009) In-vitro effects of resveratrol on trichinella spiralis. Parasitol Res 105:1139–1143
Pozio E (2019) Trichinella and trichinellosis in Europe. Vet Glas 73(2):65–84
Rowayshed G, Salama A, Abul-Fadl M, AkilaHamza S, Emad A (2013) Nutritional and chemical evaluation for pomegranate (Punica granatum L.) fruit peel and seeds powders by products. Middle East J Appl Sci 3(4):169–179
Saeed M, Naveed M, BiBi J, Kamboh AA, Arain MA, Shah QA, Alagawany M, El-Hack ME, Abdel-Latif MA, Yatoo MI, Tiwari R, Chakraborty S, Dhama K (2018) The promising pharmacological effects and therapeutic/medicinal applications of Punica Granatum L. (Pomegranate) as a functional food in humans and animals. Rec Pat Inflamm Allergy Drug Discov 12(1):24–38
Salhan M, Kumar B, Tiwari P, Sharma P, Sandhar HK, Gautam M (2011) Comparative anthelmintic activity of aqueous and ethanolic leaf extracts of Clitoria ternatea. Int J Drug Dev Res 3(1):62–69
Singh RP, Chidambara Murthy KN, Jayaprakasha GK (2002) Studies on the antioxidant activity of pomegranate (Punica granatum) peel and seed extracts using in vitro models. J Agric Food Chem 50(1):81–86
Srivastava M, Misra-Bhattacharya S (2015) Overcoming drug resistance for macro parasites. Fut Microbiol 10(11):1783–1789. https://doi.org/10.2217/fmb.15.73.PMid:26517758
Steel N, Faniyi AA, Rahman S, Swietlik S, Czajkowska BI, Chan BT, et al. (2019) TGFβ-activation by dendritic cells drives Th17 induction and intestinal contractility and augments the expulsion of the parasite Trichinella spiralis in mice. PLoS Pathog 15(4):e1007657. https://doi.org/10.1371/journal.ppat.1007657
Sun XM, Guo K, Hao CY, Zhan B, Huang JJ, Zhu X (2019) Trichinella spiralis Excretory-Secretory Products Stimulate Host Regulatory T Cell Differentiation through Activating Dendritic Cells. Cells 8(11):1404. https://doi.org/10.3390/cells8111404
Ullah F, Ayaz M, Sadiq A, Ullah F, Hussain I, Shahid M et al (2020) Potential role of plant extracts and phytochemicals against foodborne pathogens. Appl Sci (basel) 10(13):4597. https://doi.org/10.3390/app10134597
Vučić V, Grabež M, Trchounian A, Arsić A (2019) Composition and potential health benefits of pomegranate: a review. Curr Pharm Des 25(16):1817–1827
Wang T, Men R, Hu M, Fan X, Yang X, Huang X et al (2018) Protective effects of Punica granatum (pomegranate) peel extract on concanavalin A-induced autoimmune hepatitis in mice. Biomed Pharmacother 100:213–220. https://doi.org/10.1016/j.biopha.2017.12.110.PMid:29428670
Wang N, Bai X, Tang B, Yang Y, Wang X, Zhu H, et al (2020) Primary characterization of the immune response in pigs infected with Trichinella spiralis. Vet Res 51(1):17. https://doi.org/10.1186/s13567-020-0741-0
Yadav AK (2012) Efficacy of Lasia spinosa leaf extract in treating mice infected with Trichinella spiralis. Parasitol Res 110(1):493–498
Yones DA, Badary DM, Sayed HMB, Bayoumi SAH, Khalifa AA, El-Moghazy AM (2016) Comparative evaluation of anthelmintic activity of edible and ornamental pomegranate ethanolic extracts against Schistosoma mansoni. BioMed Res Int. https://doi.org/10.1155/2016/2872708
Zhao SS, Ma DX, Zhu Y, Zhao JH, Zhang Y, Chen JQ, Sheng ZL (2018) Antidiarrheal effect of bioactivity-guided fractions and bioactive components of pomegranate (Punica granatum L.) peels. Neurogastroenterol Motil 30(7):e13364
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The authors deeply appreciate the support given by Electron Microscope Department at Theodor Bilharz Research Institute, Giza, Egypt for imaging the electron microscopic pictures.
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El-Sayed, S.H., Mahmoud, S.S., El-Shenawy, A.M. et al. Anti-helminthic effect of Punica granatum peel extract on Trichinella spiralis worms and muscle larvae: in vitro and in vivo studies. J Parasit Dis 47, 416–424 (2023). https://doi.org/10.1007/s12639-023-01586-7
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DOI: https://doi.org/10.1007/s12639-023-01586-7