Abstract
There is an urgent need to develop natural antimicrobials for the control of rapidly mutating drug-resistant bacteria and poultry viruses. Five extracts were prepared using diethyl ether, ethyl acetate, methanol, 1-butanol and n-hexane from abdominal fats of Varanus griseus locally known as Indian desert monitor. Antibacterial, antioxidant and antiviral activities from oil extracts were done through disc diffusion method, stable 1,1-diphenyl-2-picrylhydrazyl (DPPH) free radical scavenging assay and in ovo antiviral assay, respectively. The gas chromatography mass spectrometry (GC–MS) analyses were used to determine principal active compounds and chemical profile of each oil extract. n-Hexane extract showed clear zones of inhibition (ZOI) against Staphylococcus aureus, Escherichia coli and Klebsiella pneumoniae (12 ± 0.5 mm, 9 ± 0.5 mm, and 9 ± 0.5 mm) while diethyl ether extract exhibited significant antibacterial activity (11 ± 0.5 mm) against Proteus vulgaris only. In case of drug-resistant strains, methanol extract was active (6 ± 0.5 mm) against Staphylococcus aureus, whereas n-hexane extract has shown ZOI 11 ± 0.5 mm against P. aeruginosa. Range of percentage scavenging activity of V. griseus oil extracts from DPPH free radical assay was 34.9–70.7%. For antiviral potential, growth of new castle disease virus (NDV) was effectively inhibited by all five extracts (HA titer = 0–4). The highest antiviral activity against avian influenza virus (H9N2) was observed from methanol, diethyl ether and 1-Butanol oil extracts with HA titers of 2, 2 and 0, respectively. Methanol, diethyl ether, 1-butanol and n-hexane oil extracts produced best hemagglutination assay (HA) titer values (0, 0, 4 and 0) against infectious bronchitis virus (IBV). Ethyl acetate and 1-Butanol extract exhibited good antiviral potential against infectious bursal disease virus (IBDV) with indirect hemagglutination assay (IHA) titers of 8 and 4, respectively. Main classes of identified compounds through gas chromatography were aldehydes, fatty acids, phenols and esters. GC–MS identified 11 bioactive compounds in V. griseus oil extracts. It is summarized that V. griseus oil has strong antioxidant activity and good antimicrobial potential because of its bioactive compounds.
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Abbreviations
- DPPH:
-
1,1-Diphenyl-2-picrylhydrazyl
- GC–MS:
-
Gas chromatography mass spectrometry
- NDV:
-
Newcastle disease virus
- AIV:
-
Avian influenza virus
- IBV:
-
Infectious bronchitis virus
- IBDV:
-
Infectious bursal disease virus
- HA:
-
Hemagglutination test
- IHA:
-
Indirect hemagglutination test
- ZOI:
-
Zone of inhibition
- INT:
-
P-iodonitro-phenyltetrazolium violet
- MIC:
-
Minimum inhibition concentration
- PBS:
-
Phosphate buffer saline solution
- RPM:
-
Revolutions per minute
- PI:
-
Post-infection
- EDTA:
-
Ethylenediaminetetraacetic acid
References
Abubakar MN, Majinda RRT (2016) GC-MS Analysis and preliminary antimicrobial activity of Albizia adianthifolia (Schumach) and Pterocarpus angolensis (DC). Medicines 3:3. https://doi.org/10.3390/MEDICINES3010003
Ahmad S, Akram M, Riaz M et al (2021) Zootherapy as traditional therapeutic strategy in the Cholistan desert of Bahawalpur-Pakistan. Vet Med Sci. https://doi.org/10.1002/vms3.491
Ak T, Gülçin I (2008) Antioxidant and radical scavenging properties of curcumin. Chem Biol Interact 174:27–37. https://doi.org/10.1016/j.cbi.2008.05.003
Akhtar S, Nazar B, Shariq A (2020) The comparison of sensitivity and specificity of ELISA-based microneutralization test with hemagglutination inhibition test to evaluate neutralizing antibody against influenza virus (H1N1). J Med Physiol Biophys. https://doi.org/10.7176/JMPB/65-08
Alkadi H (2018) A review on free radicals and antioxidants. Infect Disord Drug Targets 20:16–26. https://doi.org/10.2174/1871526518666180628124323
Altaf M, Abbasi AM, Umair M et al (2020) The use of fish and herptiles in traditional folk therapies in three districts of Chenab riverine area in Punjab Pakistan. J Ethnobiol Ethnomed. https://doi.org/10.1186/S13002-020-00379-Z
Alves RR, Alves HN (2011) The faunal drugstore: animal-based remedies used in traditional medicines in Latin America. J Ethnobiol Ethnomed 7:9. https://doi.org/10.1186/1746-4269-7-9
Alves RRN, Rosa IL (2007) Zootherapy goes to town: the use of animal-based remedies in urban areas of NE and N Brazil. J Ethnopharmacol 113:541–555. https://doi.org/10.1016/J.JEP.2007.07.015
Alves RRN, Rosa IL, Santana GG (2007) The role of animal-derived remedies as complementary medicine in Brazil. Bioscience 57:949–955. https://doi.org/10.1641/B571107
Alves RRN, Barbosa JAA, Santos SLDX et al (2011) Animal-based remedies as complementary medicines in the semi-arid region of Northeastern Brazil. Evid Based Complement Altern Med 2011:1–15. https://doi.org/10.1093/ecam/nep134
Alves E, Dias M, Lopes D et al (2020) Antimicrobial lipids from plants and marine organisms: an overview of the current state-of-the- art and future prospects. Antibiotics 9:1–88
Arshad M, Ruby T, Shahzad MI et al (2022) An antimicrobial activity of oil extracted from Saara hardwickii. Braz J Biol 84:1–7. https://doi.org/10.1590/1519-6984.253508
Bajpai SK, Chandra K (1990) Studies on the antiviral properties of plants with special reference to Zingiber capitatum. Fitoterapia 61:3–8
Balouiri M, Sadiki M, Ibnsouda SK (2016) Methods for In vitro evaluating antimicrobial activity: a review. J Pharm Anal 6:71–79. https://doi.org/10.1016/j.jpha.2015.11.005
Balta I, Stef L, Pet I et al (2020) Antiviral activity of a novel mixture of natural antimicrobials, In vitro, and in a chicken infection model In vivo. Sci Rep 10:1–10. https://doi.org/10.1038/s41598-020-73916-1
Barbour EK, Yaghi RH, Jaber LS et al (2008) Safety and antiviral activity of essential oil against avian Influenza and NewCastle disease viruses. Int J Appl Res Vet Med 8:60–64
Bennett DC, Code WE, Godin DV, Cheng KM (2008) Comparison of the antioxidant properties of emu oil with other avian oils. Aust J Exp Agric 48:1345. https://doi.org/10.1071/EA08134
Cai Y, Luo Q, Sun M, Corke H (2004) Antioxidant activity and phenolic compounds of 112 traditional Chinese medicinal plants associated with anticancer. Life Sci 74:2157–2184. https://doi.org/10.1016/J.LFS.2003.09.047
Churchward CP, Alany RG, Snyder LAS (2018) Alternative antimicrobials: the properties of fatty acids and monoglycerides. Crit Rev Microbiol 44:561–570
Cunha-Neto A, Carvalho LA, Castro VS et al (2020) Salmonella anatum, S. infantis and S. schwarzengrund in Brazilian cheeses: occurrence and antibiotic resistance profiles. Int J Dairy Technol 73:296–300. https://doi.org/10.1111/1471-0307.12636
da Brazil MVS, de Souza MB, Ferraz VP et al (2020) Antibacterial, total phenols, antioxidant, and fatty acids of the lyophilized body fat of Podocnemis expansa (Schweigger, 1812) from farm in Acre State, Brazil. J Med Plants Res 14:458–467. https://doi.org/10.5897/jmpr2020.6995
Danish MT, Aslam HMM, Dur-E Z (2018) The comparison of sensitivity and specificity of ELISA-based microneutralization test with hemagglutination inhibition test to evaluate neutralizing antibody against influenza virus (H1N1). Pakistan J Med Heal Sci 12:1052–1054
de Dias DQ, Sales DL, Andrade JC et al (2019) Antibacterial and antibiotic modifying activity evaluation of ruminants’ body fat used as zootherapeutics in ethnoveterinary practices in Northeast Brazil. J Ethnopharmacol 233:87–93. https://doi.org/10.1016/j.jep.2018.12.012
Dias DDQ, Cabral MES, Sales DL et al (2013) Chemical composition and validation of the ethnopharmacological reported antimicrobial activity of the body fat of Phrynops geoffroanus used in traditional medicine. Evid Based Complement Altern Med. https://doi.org/10.1155/2013/715040
Fernando SA, Pang S, McKew GL et al (2020) Evaluation of the Haemophilus influenzae EUCAST and CLSI disc diffusion methods to recognize aminopenicillin and amoxicillin/clavulanate resistance. J Antimicrob Chemother 75:2594–2598. https://doi.org/10.1093/jac/dkaa229
Ferreira FS, Brito SV, Ribeiro SC et al (2009) Animal-based folk remedies sold in public markets in Crato and Juazeiro do Norte, Ceará Brazil. BMC Complement Altern Med 9:17. https://doi.org/10.1186/1472-6882-9-17
Ferreira FS, Brito SV, Saraiva RA et al (2010) Topical anti-inflammatory activity of body fat from the lizard Tupinambis merianae. J Ethnopharmacol 130:514–520. https://doi.org/10.1016/j.jep.2010.05.041
Ferreira FS, Brito SV, Coutinho HDM et al (2018) Vertebrates as a bactericidal agent. EcoHealth 15:619–626. https://doi.org/10.1007/s10393-018-1345-2
Hashmi MUA, Khan MZ, Amtyaz NU, Ul Huda N (2013) Current status, distribution and threats of Varanus species (Varanus bengalensis & Varanus griseus) in Karachi & Thatta of Sindh. Int J Fauna Biol Stud 1:34–38
Hayashi K, Kamiya M, Hayashi T (1995) Virucidal effects of the steam distillate from Houttuynia cordata and its components on HSV-1, influenza virus, and HIV. Planta Med 61:237–241. https://doi.org/10.1055/S-2006-958063
Hussain MA, Gorsi MS (2004) Antimicrobial activity of Nerium oleander linn. Asian J Plant Sci 3:177–180. https://doi.org/10.3923/ajps.2004.177.180
Hussain T, Fatima I, Rafay M et al (2015) Comparison of antibacterial potential from leaves and fruits of different herbs and shrubs of family solanaceae. Int J Agric Biol 17:1249–1254. https://doi.org/10.17957/IJAB/15.0039
Inguglia L, Chiaramonte M, Di Stefano V et al (2020) Salmo salar fish waste oil: fatty acids composition and antibacterial activity. PeerJ. https://doi.org/10.7717/peerj.9299
Jiang L, Wang F, Han F et al (2013) Evaluation of diffusion and dilution methods to determine the antimicrobial activity of water-soluble chitosan derivatives. J Appl Microbiol 114:956–963. https://doi.org/10.1111/jam.12111
Kakati LN, Ao B, Doulo V (2006) Indigenous knowledge of zootherapeutic use of vertebrate origin by the Ao tribe of Nagaland. J Hum Ecol 19:163–167. https://doi.org/10.1080/09709274.2006.11905874
Khunsap S (2016) Antioxidant, anticancer cell lines and physiochemical evaluation of cobra oil. Int J Pure Appl Biosci 4:21–27. https://doi.org/10.18782/2320-7051.2296
Kim YG, Lee JH, Raorane CJ et al (2018) Herring oil and omega fatty acids inhibit Staphylococcus aureus biofilm formation and virulence. Front Microbiol. https://doi.org/10.3389/FMICB.2018.01241
Lobo V, Patil A, Phatak A, Chandra N (2010) Free radicals, antioxidants and functional foods: impact on human health. Pharmacogn Rev 4:118–126
Mishra B, Akhila MVA, Thomas A et al (2020) Formulated therapeutic products of animal fats and oils: future prospects of zootherapy. Int J Pharm Investig 10:112–116. https://doi.org/10.5530/ijpi.2020.2.20
Okwor EC, Eze DC, Okonkwo KE (2011) Serum antibody levels against infectious bursal disease (IBD) virus in village chickens using indirect haemagglutination (IHA) test. J Anim Vet Adv 10:2683–2686. https://doi.org/10.3923/javaa.2011.2683.2686
Oliveira OP, Sales DL, Dias DQ et al (2014) Antimicrobial activity and chemical composition of fixed oil extracted from the body fat of the snake Spilotes pullatus. Pharm Biol 52:740–744. https://doi.org/10.3109/13880209.2013.868495
Purkait S, Bhattacharya A, Bag A, Chattopadhyay RR (2020) Synergistic antibacterial, antifungal and antioxidant efficacy of cinnamon and clove essential oils in combination. Arch Microbiol 202:1439–1448. https://doi.org/10.1007/s00203-020-01858-3
Rahman MM, Ahmad SH, Mohamed MTM, Ab Rahman MZ (2014) Antimicrobial compounds from leaf extracts of Jatropha curcas, Psidium guajava, and Andrographis paniculata. Sci World J. https://doi.org/10.1155/2014/635240
Rajbhandari M, Wegner U, Jülich M et al (2001) Screening of nepalese medicinal plants for antiviral activity. J Ethnopharmacol 74:251–255. https://doi.org/10.1016/S0378-8741(00)00374-3
Reichling J, Schnitzler P, Suschke U, Saller R (2009) Essential oils of aromatic plants with antibacterial, antifungal, antiviral, and cytotoxic properties–an overview. Forsch Komplementmed 16:79–90. https://doi.org/10.1159/000207196
Riós-Orjuela JC, Falcón-Espitia N, Arias-Escobar A et al (2020) Knowledge and interactions of the local community with the herpetofauna in the forest reserve of Quininí (Tibacuy-Cundinamarca, Colombia). J Ethnobiol Ethnomed 16:1–11. https://doi.org/10.1186/s13002-020-00370-8
Sales DL, Oliveira OP, Cabral MES et al (2015) Chemical identification and evaluation of the antimicrobial activity of fixed oil extracted from Rhinella jimi. Pharm Biol 53:98–103. https://doi.org/10.3109/13880209.2014.911331
Schnitzler P, Astani A, Reichling J (2011) Screening for antiviral activities of isolated compounds from essential oils. Evid Based Complement Altern Med. https://doi.org/10.1093/ECAM/NEP187
Setzer WN (2016) Essential oils as complementary and alternative medicines for the treatment of influenza. Am J Essent Oils Nat Prod. 4:16–22
Shams WA, Rehman G, Onoja SO et al (2019) In vitro antidiabetic, anti-inflammatory and antioxidant potential of the ethanol extract of Uromastyx hardwickii skin. Trop J Pharm Res 18:2109–2115. https://doi.org/10.4314/tjpr.v18i10.16
Simitzis PE (2017) Enrichment of animal diets with essential oils—a great perspective on improving animal performance and quality characteristics of the derived products. Med 4:35. https://doi.org/10.3390/MEDICINES4020035
Spencer PVD, Libardi SH, Dias FFG et al (2021) Chemical composition, antioxidant and antibacterial activities of essential oil from Cymbopogon densiflorus (Steud.) stapf flowers. J Essent Oil-Bearing Plants 24:40–52. https://doi.org/10.1080/0972060X.2020.1862711
Tanhaeian A, Sekhavati MH, Moghaddam M (2020) Antimicrobial activity of some plant essential oils and an antimicrobial-peptide against some clinically isolated pathogens. Chem Biol Technol Agric 7:13. https://doi.org/10.1186/s40538-020-00181-9
Thormar H, Isaacs CE, Brown HR et al (1987) Inactivation of enveloped viruses and killing of cells by fatty acids and monoglycerides. Antimicrob Agents Chemother 31:27–31. https://doi.org/10.1128/AAC.31.1.27
Thirupathi K, Chandrakala G, Krishna L et al (2018) Antibacterial activity of skin secretion and its extraction from the toad Bufo melanostictus. Eur J Pharm Med Res 3:283–286
Vijayan P, Raghu C, Ashok G et al (2004) Antiviral activity of medicinal plants of Nilgiris. Indian J Med Res 120:24–29
Yaqeen Z, Naqvi N-H, Sohail T et al (2013) Screening of solvent dependent antibacterial activity of Prunus domestica. Pak J Pharm Sci 26(2):409–414. https://www.researchgate.net/publication/235776249
Zamora-Gasga VM, Loarca-Piña G, Vázquez-Landaverde PA et al (2015) In vitro colonic fermentation of food ingredients isolated from Agave tequilana Weber var. azul applied on granola bars. Lwt 60:766–772. https://doi.org/10.1016/j.lwt.2014.10.032
Zheng W, Wang SY (2001) Antioxidant activity and phenolic compounds in selected herbs. J Agric Food Chem 49:5165–5170. https://doi.org/10.1021/jf010697n
Acknowledgements
The authors would like to thank “Institute of Pure and Applied Biology” Bahauddin Zakariya University, Multan, Pakistan for facilitating to conduct this study and Laboratorio de Biotecnología Farmacéutica, Centro de Biotecnología Genómica, Instituto Politécnico Nacional, Reynosa, Mexico for GC–MS analysis.
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AAK and TR: designed the study and supervised the experiments, SA: collected and verified animal specimens, SA and MIS: performed the laboratory experiments. GR and DVNC: executed chromatographic analysis of oil, and SA: corrected and finalized the manuscript.
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Communicated by Erko Stackebrandt.
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Ahmad, S., Ruby, T., Shahzad, M.I. et al. Antimicrobial, antioxidant, antiviral activity, and gas chromatographic analysis of Varanus griseus oil extracts. Arch Microbiol 204, 531 (2022). https://doi.org/10.1007/s00203-022-03138-8
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DOI: https://doi.org/10.1007/s00203-022-03138-8