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. 2024 Jan 1;25(1):565. doi: 10.3390/ijms25010565

Table 8.

Antimicrobial, larvicidal, and insecticidal activity of essential oil and extracts of G. procumbens.

Essential Oil/Extract Microorganism/Activity Results Positive Control References
antibacterial activity
essential oil from herbs (aerial parts) microdilution method (%):
Acinetobacter baumannii (MIC = 0.25%, v/v),
Aeromonas sobria (0.25%),
Enterococcus faecalis (>2.0%),
Escherichia coli (0.5%),
Klebsiella pneumoniae (1.0%),
Pseudomonas aeruginosa (>2.0%),
Salmonella typhimurium (0.5%),
Serratia marcescens (0.5%),
Staphylococcus aureus (2.0%)		 − [145]
disc diffusion method; zone of inhibition diameter (mm):
Staphylococcus aureus: 19.5 mm (essential oil concentration: 0.1%, v/v), 20.4 mm (0.5%), 22.0 mm (1.0%),
Escherichia coli: 18.2 mm (0.1%), 18.8 mm (0.5%), 19.9 mm (1.0%)		 chloramphenicol (0.1 mg/mL):
S. aureus: 23.2 mm,
E. coli: 21.9 mm		 [49]
essential oil from leaves disc diffusion method; zone of inhibition diameter (mm):
Staphylococcus aureus (7.33 mm),
Staphylococcus capitis (8.33 mm),
Staphylococcus epidermidis (4.33–5.67 mm),
Staphylococcus haemoliticus (4.33–6.33 mm),
Staphylococcus hominis (3.33–5.67 mm)
microdilution method (μL/mL):
Staphylococcus aureus (MIC = 12.50 μL/mL),
Staphylococcus capitis (12.50 μL/mL),
Staphylococcus epidermidis (25.00 μL/mL),
Staphylococcus haemoliticus (12.50–25.00 μL/mL),
Staphylococcus hominis (25.00 μL/mL)		 − [44]
microdilution method (mg/mL):
Escherichia coli (MIC = 6.33 mg/mL),
Aeromonas caviae (3.16 mg/mL),
Serratia marcescens (6.33 mg/mL),
Staphylococcus aureus (6.33 mg/mL),
Streptococcus pneumoniae (12.67 mg/mL),
Candida albicans (1.82 mg/mL),
Candida tropicalis (7.29 mg/mL),
Candida glabrata (7.29 mg/mL),
Mycobacterium fortuitum (3.64 mg/mL),
Mycobacterium abscessus (3.64 mg/mL)		 − [42]
disc diffusion method; zone of inhibition diameter (mm):
Staphylococcus aureus (23.9 mm),
Escherichia coli (7.9 mm)		 amoxicillin (50 μg/mL):
S. aureus (33.8 mm),
E. coli (19.2 mm)		 [47]
microdilution method (MIC and MBC in % essential oil):
Pasteurella multocida (MIC = 0.5%; MBC not possible to determine);
disc diffusion method: median inhibition zone radius (mm); interquartile range:
Pasteurella multocida (14.5 mm; 1.25);
Mannheimia haemolytica (15.5 mm);
Mannheimia clade (12 mm)		 growth control (with only sterile blank filter paper discs); negative control (wintergreen oil without bacterial inoculum) [48]
microdilution method (mg/mL):
Staphylococcus aureus (MIC = 14.0 mg/mL),
Enterococus faecalis (16.4 mg/mL),
Escherichia coli (9.4 mg/mL),
Acinetobacter baumanii (8.2 mg/mL)		 gentamicin:
S. aureus (MIC = 0.016 mg/mL),
E. faecalis (0.064 mg/mL),
E. coli (0.004 mg/mL),
A. baumanii (0.032 mg/mL)		 [10]
clinical strains and environmental bacteria
microdilution method (mg/mL):
Staphylococcus aureus (pus, MIC = 14.1 mg/mL),
Staphylococcus aureus (pharynx, 14.1 mg/mL),
Enterococcus faecalis (urine, 16.7 mg/mL),
Enterococcus faecalis (respiratory, 16.1 mg/mL),
Escherichia coli (wound, 10.0 mg/mL),
Escherichia coli (bronchia, 8.8 mg/mL),
Acinetobacter baumanii (sputum, 8.5 mg/mL),
Acinetobacter baumanii (sink, 8.5 mg/mL)		 −
essential oil from fruits microdilution method (mg/mL):
Staphylococcus aureus (MIC = 13.5 mg/mL),
Enterococus faecalis (15.3 mg/mL),
Escherichia coli (8.8 mg/mL),
Acinetobacter baumanii (8.2 mg/mL)		 gentamicin:
S. aureus (MIC = 0.016 mg/mL),
E. faecalis (0.064 mg/mL),
E. coli (0.004 mg/mL),
A. baumanii (0.032 mg/mL)
clinical strains and environmental bacteria;
microdilution method (mg/mL):
Staphylococcus aureus (pus, MIC = 14.1 mg/mL),
Staphylococcus aureus (pharynx, 13.2 mg/mL),
Enterococcus faecalis (urine, 15.5 mg/mL),
Enterococcus faecalis (respiratory, 16.4 mg/mL),
Escherichia coli (wound, 9.7 mg/mL),
Escherichia coli (bronchia, 8.8 mg/mL),
Acinetobacter baumanii (sputum, 8.8 mg/mL),
Acinetobacter baumanii (sink, 8.5 mg/mL)		 −
commercial essential oil microdilution method (mg/mL):
Listeria monocytogenes (MIC = 0.29 mg/mL),
Staphylococcus aureus (0.36 mg/mL),
Escherichia coli (0.36 mg/mL),
Pseudomonas aeruginosa (0.18 mg/mL),
Salmonella typhimurium (0.36 mg/mL)		 streptomycin:
L. monocytogenes (MIC = 0.05 mg/mL),
S. aureus (0.05 mg/mL),
E. coli (0.20 mg/mL),
P. aeruginosa (0.05 mg/mL),
S. typhimurium (0.10 mg/mL)		 [9]
growth (%) of bacteria in the tested sample
(milk + bacteria + essential oil at a concentration 4%, v/v):
Staphylococcus aureus: 108%
Staphylococcus chromogenes: 65%
Streptococcus uberis: 159%		 control:
sample containing milk + bacteria:
100% bacterial growth		 [144]
Pseudominas tolaasii (MIC = 0.08 µg/mL; MBC = 0.16 µg/mL),
Clavibacter michiganensis subsp. michiganensis (MIC = 0.08 µg/mL; MBC = 0.16 µg/mL),
Xanthomonas campestris pv. phaseoli (MIC = 0.02 µg/mL; MBC = 0.02 µg/mL)		 − [53]
microdilution method (μL/mL):
Aerococcus spp. (MIC = 12.5 μL/mL); Aerococcus viridans (6.25 μL/mL); Aeromonas spp. (3.12 μL/mL); Aeromonas bestiarum (6.25 μL/mL); Aeromonas salmonicida (3.12 μL/mL); Escherichia vulgaris (25.0 μL/mL); Enterococcus faecium (3.12 μL/mL); Enterococcus moravensis (3.12 μL/mL); Enterococcus aquimarinus (3.12 μL/mL); Pseudomonas fluorescens (25.0 μL/mL); Pseudomonas frederiksbergensis (25.0 μL/mL); Pseudomonas gessardii (25.0 μL/mL); Pseudomonas ludensis (25.0 μL/mL); Pseudomonas proteolitica (6.25 μL/mL); Shewanella baltica (6.25 μL/mL); Yersinia enterocolitica (6.25 μL/mL); Yersinia ruckeri (6.25 μL/mL); Yersinia spp. (6.25 μL/mL); Vagococcus spp. (6.25 μL/mL)		 − [55]
ethanol–water (55:45, v/v) extract from leaves Neisseria gonorrhoeaeMIC: >512 µg/mL penicillin G:
MIC = 128 µg/mL		 [148]
alcoholic extract from herb (aerial parts) concentration 5% (w/v)
disc diffusion method; zone of inhibition diameter (mm):
Staphylococcus aureus: 20.8 mm,
Escherichia coli: 19.0 mm		 chloramphenicol (0.1 mg/mL):
S. aureus: 23.2 mm,
E. coli: 21.9 mm		 [49]
antifungal activity
essential oil from herb (aerial parts) Candida albicans (MIC = 0.25%, v/v) − [145]
disc diffusion method; zone of inhibition diameter (mm):
Candida albicans: 16.5 mm (essential oil concentration 0.1%, v/v), 17.2 mm (0.5%), 18.0 mm (1.0%);
Aspergillus niger: 15.9 mm (0.1%), 16.6 mm (0.5%), 17.2 mm (1.0%)		 ketoconazole (0.1 mg/mL):
C. albicans (18.8 mm),
A. niger (18.1 mm)		 [49]
essential oil from leaves disc diffusion method; zone of inhibition diameter (mm):
Torulaspora delbruecki (32.66 mm),
Debaromyces hanseni spp. hanseni (29.65 mm),
Kluyveromyces maxianus spp. lactis (36.54 mm),
Pichia polymorpha (40.23 mm),
Candida lactis spp. condensi (14.23 mm),
Candida humulis (16.74 mm),
Saccharomyces cerevisiae (16.90 mm),
Zygosaccharomyces lactis (9.67 mm),
Zygosaccharomyces pombe (8.80 mm),
Torula lactis (15.81 mm)
microdilution method (μg/mL):
Torulaspora delbruecki (MIC = 8.79 μg/mL; MFC = 5.86 μg/mL),
Debaromyces hanseni spp. hanseni (17.58 μg/mL; 11.72 μg/mL),
Kluyveromyces maxianus spp. lactis (5.85 μg/mL; 2.93 μg/mL),
Pichia polymorpha (4.39 μg/mL; 19.53 μg/mL),
Candida lactis spp. condensi (29.30 μg/mL; 17.58 μg/mL),
Candida humulis (35.16 μg/mL; 23.44 μg/mL),
Saccharomyces cerevisiae (58.60 μg/mL; 35.16 μg/mL),
Zygosaccharomyces lactis (562.50 μg/mL; 78.13 μg/mL),
Zygosaccharomyces pombe (375.00 μg/mL; 156.25 μg/mL),
Torula lactis (234.38 μg/mL; 78.13 μg/mL)		 − [43]
commercial essential oil microdilution method (mg/mL):
Aspergillus flavus (MIC = 2.92 mg/mL),
Aspergillus fumigatus (0.73 mg/mL),
Aspergillus niger (0.73 mg/mL),
Aspergillus ochraceus (1.46 mg/mL),
Penicillium funiculosum (0.73 mg/mL),
Penicillium ocrochloron (1.46 mg/mL)		 ketoconazole:
A. flavus (1.50 mg/mL),
A. fumigatus (0.20 mg/mL),
A. niger (0.20 mg/mL),
A. ochraceus (0.20 mg/mL),
P. funiculosum (0.20 mg/mL),
P. ocrochloron (1.00 mg/mL)		 [9]
disc diffusion method; zone of inhibition diameter (cm):
Phomopsis azadirachtae: 7.00 cm (essential oil concentration 500 ppm), 5.40 cm (1000 ppm), 0.00 cm (1500 ppm), 0.00 cm (2000 ppm), 0.00 cm (2500 ppm)		 Tween 20:
9.00 cm		 [146]
antifungal activity against 11 plant pathogens − [147]
Trichophyton mentagrophytes
disc diffusion method; zone of inhibition diameter (mm): 0 mm
box vapour assay: MFD > 100 μg/mL		 − [52]
Fusarium, including F. verticilliodies, F. napiforme, and F. delphinoides
MIC: 2400–38,400 µg/mL		 natamycin:
MIC: 2–256 µg/mL		 [51]
alcoholic extract from herb (aerial parts) concentration 5% (w/v)
disc diffusion method; zone of inhibition diameter (mm):
Candida albicans: 17.0 mm,
Aspergillus niger: 16.8 mm		 ketoconazole (0.1 mg/mL):
C. albicans (18.8 mm),
A. niger (18.1 mm)		 [49]
ethanol–water (1:1, v/v) extract from leaves disc diffusion method; zone of inhibition diameter (mm):
Microsporum gypseum: 7.9 mm,
Trichophyton mentagrophytes: 7.7 mm;
the extract did not work on: Saccharomyces cerevisiae, Cryptococcus neoformans, Candida albicans, and Aspergillus fumigatus 		berberine (2 mg):
M. gypseum: 18.0 mm,
T. mentagrophytes: 20.0 mm,
S. cerevisiae: 10.0 mm,
C. neoformans: 26.0 mm,
C. albicans: 16.0 mm,
A. fumigatus: 10.0 mm		 [149]
larvicidal, insecticidal, and other activity
essential oil from leaves Cortaderia selloana (pampas grass):
in vitro effect against seed germination: 83% (concentration of wintergreen oil 0.125 μL/mL), 77% (0.25 μL/mL), 56% (0.5 μL/mL), 19% (1 μL/mL);
Nicotiana glauca (tree tobacco):
in vitro effect against seed germination: 90% (concentration of wintergreen oil 0.125 μL/mL), 87% (0.25 μL/mL), 91% (0.5 μL/mL), 79% (1 μL/mL)		 control:
untreated seedlings
C. selloana: 84%;
N. glauca: 94%		 [45]
influenza A/WS/33 virus
lack of anti-influenza activity (no reduction in visible cytopathic effects of influenza A/WS/33 virus activity) at a concentration of 100 μg/mL		 control:
oseltamivir (100 μg/mL): 49%		 [150]
Sitophilus oryzae (rice weevil) and Rhyzopertha dominica (lesser grain borer)
method: jars with wheat seeds, paper discs treated with wintergreen oil (without any solvents), and 50 adults (4–6 days old) of both insects;
antifeedant activity:
S. oryzae: % seed damage (0.0%), % weight loss (0.0%), FDI (100%),
R. dominica: % seed damage (0.0%), % weight loss (0.0%), FDI (100%);
fumigant toxicity:
S. oryzae: LC50 = 58.62 μL/L (51–85–64.47 μL/L),
R. dominica: LC50 = 2.71 μL/L (2.36–3.03 μL/L)		 control:
paper discs without wintergreen oil and any solvents; antifeedant activity:
S. oryzae: % seed damage (65.04%), % weight loss (8.26%), FDI (0%);
R. dominica: % seed damage (76.04%), % weight loss (5.33%), FDI (0%)		 [39]
Callosobruchus chinensis (pulse beetle, Chinese bruchid, cowpea bruchid)
method: glass cylinder with paper discs treated with wintergreen oil dissolved in acetone;
fumigant toxicity after 24 h: LC50 = 3.14 mg/L		 control:
paper discs treated with acetone		 [151]
commercial essential oil fourth instar larvae of Agrotis ipsilon (dark sword-grass moth)
method: antifeedant activity; Petri dishes with fresh potato leaf discs treated with wintergreen oil dissolved in water and one 2 h pre-starved insect;
method: insecticidal activity; round plastic trough with fresh potato leaf discs treated with wintergreen oil dissolved in water and 10 2 h pre-starved insects;
antifeedant activity: 19.79% (wintergreen oil concentration 0.25%), 28.41% (0.5%), 54.66% (1.0%), 87.21% (2.0%);
insecticidal activity: 2.70% (0.25%), 18.41% (0.5%), 53.64% (1.0%), 86.92% (2.0%)		 control:
leaf discs treated with water;
antifeedant activity: 4.22%,
insecticidal activity: 1.20%		 [152]
third instar larvae of Camptomyia corticalis (Cecidomyiidae gall midge, mosquito)
method: Petri dishes with paper discs treated with wintergreen oil dissolved in ethanol and 20 insects;
% mortality after 24 h: 100% (wintergreen oil concentration 1.41 mg/cm3), 97% (1.05 mg/cm3);
fumigant toxicity during a 24 h exposure: LC50 = 0.74 mg/cm3 (0.65–0.87 mg/cm3)		 control:
paper discs treated with ethanol;
positive control:
dichlorvos—conventional insecticide
fumigant toxicity: LC50 = 0.027 mg/cm3 (0.023–0.032 mg/cm3)		 [153]
fourth instar larvae of Tribolium castaneum (red flour beetle)
method: Petri dishes with paper discs treated with wintergreen oil dissolved in water and 10 insects;
repellent activity: 21.2% (wintergreen oil concentration 5 μL/mL), 43.04% (10 μL/mL), 65.96% (15 μL/mL), 80.2% (20 μL/mL);
larvicidal activity after 96 h: 18.4% (5 μL/mL), 37.78% (10 μL/mL), 54.64% (15 μL/mL), 69.91% (20 μL/mL)		 control:
paper discs treated with water;
repellent activity: 0.0%;
larvicidal activity after 24 h: 0.0% (not tested after 96 h)		 [154]
Callosobruchus maculatus (cowpea weevil, cowpea seed beetle)
method: fumigant toxicity; airtight plastic jars with 20 cowpea seeds, paper discs treated with wintergreen oil dissolved in acetone, and four newly emerged C. maculatus;
fumigant toxicity: wintergreen oil concentration 5 μL/mL, mortality after 6 h (10%), after 12 h (60%), after 24 h (85%), and 48 h (93.33%); wintergreen oil concentration 10 μL/mL, mortality after 6 h (15%), after 12 h (70%), after 24 h (90%), and 48 h (100%); wintergreen oil concentration 20 μL/mL, mortality after 6 h (45%), after 12 h (75%), after 24 h (100%), and 48 h (100%); wintergreen oil concentration 40 μL/mL, mortality after 6 h (80%), after 12 h (100%), after 24 h (100%), and 48 h (100%)
method: ovicidal activity; plastic jars with 20 cowpea seeds, paper discs treated with wintergreen oil dissolved in acetone, and 20 eggs of C. maculatus;
ovicidal activity during 48 h of exposure: wintergreen oil concentration 5 μL/mL (51.16%), wintergreen oil concentration 10 μL/mL (63.95%), wintergreen oil concentration 20 μL/mL (93.02%), wintergreen oil concentration 40 μL/mL (100.00%)		 control:
paper discs treated with acetone		 [155]
wasps: Vespula pensylvanica (western yellowjacket), Vespula vulgaris (common wasp), Vespula germanica (German yellowjacket), Dolichovespula maculate (black-faced hornet), Polistes dominula (European paper wasp) and Paralucia aurifera (golden paper wasp)
field experiment (traps set in nature); capture of social wasps in Rescue!®
repellent release rate: attractant (acetic acid+2-methyl-1-butanol)+essential oil (45 mg/day);
mean per trap per visit: Yellowjackets (Vespula pensylvanica, V. vulgaris, V. germanica, Dolichovespula maculata) = 2.3; Paper wasps (Polistes dominulus, Paralucia aurifer) = 2.0		 control:
repellent release rate: attractant without wintergreen oil (0 mg/day);
mean per trap per visit: Yellowjackets = 43.7, Paper wasps = 7.9		 [156]
Sitophylus granarius (wheat weevil)
method: Falcon tubes containing wheat treated with wintergreen oil dissolved in acetone and 20 insects;
mortality (%) after 24 h of exposure: wintergreen oil concentration 5% (100%), 4% (−), 3% (96%), 2% (81%), 1% (5%)		 control:
wheat treated with acetone;
mortality (%) after 24 h of exposure: 0%		 [56]
Anarsia lineatella (peach twig borer, fruit moth)
method: ULV (Ultra Low Volumes)-olfactory apparatus with peach twigs having one fruit and 2–5 leaves sprayed with wintergreen oil dissolved in water;
the number of eggs laid by females of A. lineatella: 59.8 eggs;
longevity of females of A. lineatella: 15.4 days;
the number of eggs laid per day by female fruit moths: after 5 days (6.4 eggs), 10 days (23.6 eggs), 15 days (31.2 eggs)		 control:
peach twigs sprayed with distilled water, soybean oil, and emulsifier—Tween 20;
the number of eggs laid by females of A. lineatella: 128 eggs; longevity of females of A. lineatella: 23.2 days;
the number of eggs laid per day by female fruit moths: after 5 days (7.4 eggs), 10 days (27.4 eggs), 15 days (32.6 eggs)		 [157]
three weeks Arabidopsis thaliana Col-0 plants (thale cress)
wintergreen oil sprayed on leaves: β-glucuronidase activity (~52 pKata/mg protein);
a strong expression of a selection of defence genes detected 1, 6, and 24 h after wintergreen oil treatment using a high-throughput qPCR-based microfluidic method;
induction of plant resistance by wintergreen oil (1 μL/mL) sprayed on leaves following inoculation with a conidial suspension of a GFP-expressing strain of the Arabidopsis thaliana fungal pathogen Colletotrichum higginsianum 48 h later: strong reduction (~60%) of pathogen development: Relative Fluorescence Units (~397 RFU)		 control:
wetting agent (benzo(1,2,3)-thiadiazole-7-carbothiolic acid (BTH)): β-glucuronidase activity (~0.8 pKata/mg protein); methyl salicylate: β-glucuronidase activity (~47 pKata/mg protein);
methyl salicylate: strong expression of a selection of defence genes;
wetting agent: ~985 RFU; commercial product; BION® (40 μg/mL): ~309 RFU		 [158]
Paederus fuscipes (rove beetle)
method: repellent activity; Petri dishes with half filter paper discs treated with wintergreen oil dissolved in acetone and 10 insects; fumigant toxicity; filter paper treated with wintergreen oil; contact toxicity; wintergreen oil dissolved in acetone applied topically to the dorsal thorax of the beetles;
contact activity: wintergreen oil concentration 0.08 μL/adult; corrected mortality after 1 h (40.00%), after 2 h (66.67%), after 4 h (73.33%), after 6 h (80.00%), after 8 h (82.22%);
contact toxicity: LC50 = 0.086 μL/adult (after 1 h of exposure), LC50 = 0.067 μL/adult (after 2 h of exposure), LC50 = 0.062 μL/adult (after 4 h of exposure), LC50 = 0.061 μL/adult (after 6 h of exposure), LC50 = 0.060 μL/adult (after 8 h of exposure);
fumigant activity: wintergreen oil concentration 2 μL/L air; corrected mortality after 1 h (24.44%), after 2 h (35.56%), after 4 h (46.67%), after 6 h (68.89%), after 8 h (80.00%);
fumigant toxicity: LC50 = 2.680 μL/L air (after 1 h of exposure), LC50 = 2.178 μL/L air (after 2 h of exposure), LC50 = 2.046 μL/L air (after 4 h of exposure), LC50 = 1.765 μL/L air (after 6 h of exposure), LC50 = 1.591 μL/L air (after 8 h of exposure);
repellent activity: wintergreen oil concentration 0.1 μL/cm2; repellency after 1 h (97.78%), after 2 h (−%), after 4 h (−%), after 6 h (−%), after 8 h (−%);
repellent activity: wintergreen oil concentration 0.01 μL/cm2; repellency after 1 h (30.00%), after 2 h (18.89%), after 4 h (12.22%), after 6 h (10.00%), after 8 h (4.44%)		 control:
paper discs treated with acetone (repellent activity);
glass vial with filter paper untreated with wintergreen oil (fumigant toxicity);
acetone applied topically on the beetles (contact toxicity)		 [58]
Paederus fuscipes (rove beetle)
method: wintergreen oil dissolved in acetone and acetylcholinesterase activity analysed in vitro:
P. fuscipes male adults: 13.0252 nM/min per mg—specific activity and 21.20% inhibition rate of enzyme activity (wintergreen oil concentration: 0.909 μL/L air), 11.0914 nM/min per mg—specific activity and 32.91% inhibition rate of enzyme activity (wintergreen oil concentration: 1.273 μL/L air), 10.9243 nM/min per mg—specific activity and 33.78% inhibition rate of enzyme activity (wintergreen oil concentration: 1.636 μL/L air);
P. fuscipes female adults: 17.5222 nM/min per mg—specific activity and 17.87% inhibition rate of enzyme activity (wintergreen oil concentration: 0.909 μL/L air), 13.8935 nM/min per mg—specific activity and 35.57% inhibition rate of enzyme activity (wintergreen oil concentration: 1.273 μL/L air), 12.2137 nM/min per mg—specific activity and 43.33% inhibition rate of enzyme activity (wintergreen oil concentration: 1.636 μL/L air);
method: five insects fumigated with wintergreen oil dissolved in acetone, homogenised, and acetylcholinesterase activity analysed in vivo:
P. fuscipes male adults in vivo: 17.4306 nM/min per mg—specific activity and 10.37% inhibition rate of enzyme activity (wintergreen oil concentration: 0.1 μL), 15.5895 nM/min per mg—specific activity and 19.76% inhibition rate of enzyme activity (wintergreen oil concentration: 0.5 μL), 14.9349 nM/min per mg—specific activity and 23.15% inhibition rate of enzyme activity (wintergreen oil concentration: 1.0 μL), 14.0898 nM/min per mg—specific activity and 27.50% inhibition rate of enzyme activity (wintergreen oil concentration: 2.0 μL), 13.4049 nM/min per mg—specific activity and 31.06% inhibition rate of enzyme activity (wintergreen oil concentration: 3.0 μL);
P. fuscipes female adults in vivo: 18.9940 nM/min per mg—specific activity and 20.75% inhibition rate of enzyme activity (wintergreen oil concentration: 0.1 μL), 15.8405 nM/min per mg—specific activity and 33.89% inhibition rate of enzyme activity (wintergreen oil concentration: 0.5 μL), 15.6144 nM/min per mg—specific activity and 34.87% inhibition rate of enzyme activity (wintergreen oil concentration: 1.0 μL), 15.4384 nM/min per mg—specific activity and 35.57% inhibition rate of enzyme activity (wintergreen oil concentration: 2.0 μL), 15.9878 nM/min per mg—specific activity and 33.32% inhibition rate of enzyme activity (wintergreen oil concentration: 3.0 μL)		 control:
in vitro (acetone instead of wintergreen oil):
P. fuscipes male adults: 16.5564 nM/min per mg—specific activity and 0% inhibition rate of enzyme activity;
P. fuscipes female adults: 21.5605 nM/min per mg—specific activity and 0% inhibition rate of enzyme activity;
control:
in vivo (acetone instead of wintergreen oil):
P. fuscipes male adults in vivo: 19.4385 nM/min per mg—specific activity and 0% inhibition rate of enzyme activity;
P. fuscipes female adults in vivo: 23.9783 nM/min per mg—specific activity and 0% inhibition rate of enzyme activity		 [57]
Apis mellifera (honey bees)
mortality (%) after ingestion of syrup containing wintergreen oil and ethanol (total number of bees: 1308): 1% (day 1), 24% (day 8, concentration of wintergreen oil 1000 ppm), 99% (day 14, concentration of wintergreen oil 100 000 ppm);
LD50 for wintergreen oil at 8 days exposure: 13,500 ppm;
LD50 for wintergreen oil at 14 days exposure: 800 ppm		 control:
syrup containing ethanol		 [159]
Loxosceles reclusa (brown recluse spider)
wintergreen oil presented the most significant mortality among all tested essential oils measured by direct contact or as a fumigant (inhalation) treatment for 24 h; the mortality in the fumigant toxicity test (<20%) was lower than in the contact toxicity assay		 − [160]
Schistocera gregaria (desert locust) and Locusta migratoria (migratory locust)
method: plastic boxes with wheat grass, sprayed with an emulsion containing linseed oil, 10% sodium bicarbonate solution, and wintergreen oil, and 10 locusts (half females and half males);
mortality (%): desert locust (12%), migratory locust (50%)		 control:
wheat grass sprayed with pure linseed oil or 10% sodium bicarbonate solution;
mortality (%): desert locust (0%), migratory locust (0%)		 [161]
method: Petri dishes with pre-cut filter paper squares sprayed with wintergreen oil dissolved in ethanol and 10 insects and clean green bean leaves;
mortality (%) after 1 h of exposure to the essential oil:
Orius laevigatus: wintergreen oil concentration 0.125 v/v (86.67%), 0.25 v/v (93.33%), 0.5 v/v (100%); toxicity category: 3 (moderately harmful);
Nesidiocoris tenuis: wintergreen oil concentration 0.125 v/v (86.67%), 0.25 v/v (96.67%), 0.5 v/v (100%); toxicity category: 3–4 (moderately harmful to harmful);
Macrolophus pygmaeus: wintergreen oil concentration 0.125 v/v (16.67%), 0.25 v/v (20.00%), 0.5 v/v (26.67%); toxicity category: 1 (harmless);
Encarsia formosa (chalcidoid wasp; parasitoid of greenhouse whitefly): wintergreen oil concentration 0.125 v/v (0%), 0.25 v/v (0%), 0.5 v/v (0%); toxicity category: 1 (harmless);
Eretmocerus eremicus (parasitoid of sweet potato whitefly): wintergreen oil concentration 0.125 v/v (0%), 0.25 v/v (0%), 0.5 v/v (0%); toxicity category: 1 (harmless);
mortality (%) after 3 h of exposure to the essential oil:
Orius laevigatus: wintergreen oil concentration 0.125 v/v (100%), 0.25 v/v (100%), 0.5 v/v (100%); toxicity category: 4 (harmful);
Nesidiocoris tenuis: wintergreen oil concentration 0.125 v/v (100%), 0.25 v/v (100%), 0.5 v/v (100%); toxicity category: 4 (harmful);
Macrolophus pygmaeus: wintergreen oil concentration 0.125 v/v (36.67%), 0.25 v/v (43.33%), 0.5 v/v (56.67%); toxicity category: 2 (slightly harmless);
Encarsia formosa (chalcidoid wasp; parasitoid of greenhouse whitefly): wintergreen oil concentration 0.125 v/v (3.33%), 0.25 v/v (6.67%), 0.5 v/v (10.00%); toxicity category: 1 (harmless);
Eretmocerus eremicus (parasitoid of sweet potato whitefly): wintergreen oil concentration 0.125 v/v (10.00%), 0.25 v/v (10.00%), 0.5 v/v (13.33%); toxicity category: 1 (harmless);
mortality (%) after 24 h of exposure to the essential oil:
Orius laevigatus: wintergreen oil concentration 0.125 v/v (-), 0.25 v/v (-), 0.5 v/v (-); toxicity category: -;
Nesidiocoris tenuis: wintergreen oil concentration 0.125 v/v (100%), 0.25 v/v (-), 0.5 v/v (-); toxicity category: 4 (harmful);
Macrolophus pygmaeus: wintergreen oil concentration 0.125 v/v (73.33%), 0.25 v/v (90.00%), 0.5 v/v (80.00%); toxicity category: 3–4 (moderately harmful to harmful);
Encarsia formosa (chalcidoid wasp; parasitoid of greenhouse whitefly): wintergreen oil concentration 0.125 v/v (6.67%), 0.25 v/v (13.33%), 0.5 v/v (20.00%); toxicity category: 1 (harmless);
Eretmocerus eremicus (parasitoid of sweetpotato whitefly): wintergreen oil concentration 0.125 v/v (16.67%), 0.25 v/v (23.33%), 0.5 v/v (26.67%); toxicity category: 1 (harmless)		 control:
Petri dishes with pre-cut filter paper squares sprayed with water and 10 insects and clean green bean leaves		 [163]
ethanol–water (80:20, v/v), dimethylsulfoxide (DMSO), and water extracts Leishmania mexicana (LM227 strain)
method: dye-binding method using bovine serum albumin as a standard;
% inhibition of LM227 growth per mg protein at 48 h;
ethanol–water (80:20, v/v) extract: not tested;
DMSO extract: <5% per mg protein;
water extract: <5% per mg protein		 control:
extract solvent instead of plant extract		 [162]

MIC: minimum inhibitory concentration; MBC: minimum bactericidal concentration; MFD: minimum fungicidal dose; FDI: feeding deterrent index.