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| Species | Purpose | Procedure | Reference |
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| S. venezuelensis | L3 larvae motility and viability observation | (i) 100 L3 decontaminated larvae were added to water in 96-well plates.
(ii) Compounds were added and plates were incubated at 28°C for 72 h.
(iii) Larvae viability was quantified through the XTT colorimetric method. and by monitoring their movement (registered through exposure to light for 2 min using an inverted microscope and a video camera).
(iv) Observations were made at 24, 48, and 72 hours after treatment.
(v) Larvae were considered dead when no movement was detected after 2 min of observation. | [16] |
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| S. ratti | Compounds with egg hatching effect | (i) Feces were homogenized in water and filtered through sieves with 250, 125 e 40 μm.
(ii) Eggs were collected and centrifuged (5 min, 3,500 ×g).
(iii) Supernatant was removed and NaCl solution (350 g/L) was added.
(iv) The mixture was homogenized and then centrifuged (5 min at 3,500 ×g). | [22] |
(v) The supernatant was filtered through a 40 μm sieve and washed with water for egg extraction.
(vi) Eggs were distributed in 96-well plates (100 eggs per well, 200 μL final vol.).
(vii) Plates were incubated at 23°C for 48 h.
(viii) Incubation was stopped and formaldehyde solution (10% in PBS) was added.
(ix) L1 larvae and eggs were counted from 200 μL aliquots and hatched percentage was calculated. |
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| S. ratti | Compounds affecting L3 larvae migration | (i) L3 larvae were incubated for 3 h at 20°C in PBS solution mixed with the extracts.
(ii) Larvae were washed 3 times in PBS solution and centrifuged.
(iii) 800 μL of larvae suspension at a concentration of 1000 L3/mL was placed over a 20 μm filter.
(iv) The filter was added to a conic tube in contact with the PBS solution surface.
(v) After 3 h, larvae above the filter were discarded and the ones that actively penetrated through the mesh to the PBS were counted using optical microscope and the migration percentage was calculated. | [23] |
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| S. ratti | Compounds with larvicidal activity in L3 larvae | (i) L3 larvae were washed 3 times in PBS and incubated in 6-well plates containing 4 mL PBS solution.
(ii) Compounds were added and larvae were incubated for 96 h (25°C, 5% CO2).
(iii) Observations were made at 1, 2, 24, 48, 72, and 96 h using optical microscope. | [17] |
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| S. stercoralis | Drugs affecting L3 larvae motility | (i) ~1 g of feces was smeared at the center of a piece of filter paper and added in a glass tube. 5 mL of the tested drug was added to the bottom of the tube incubated at 25°C.
(ii) The tubes were analyzed at the 3rd and 5th days to assess drug effect on worm viability through their motility. | [24] |
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| S. stercoralis | Compounds affecting L3 larvae mortality | (i) Dog feces were added to agar plates and left at room temperature for 3 to 5 days.
(ii) L3 larvae that migrated to the plate lid were collected, washed in saline solution (0,85%, pH 7.4), and centrifuged (2000 rpm, 5 min).
(iii) Pellet was washed 3 times in PBS (0.1 M, pH 7.4) with penicillin G (1000 IU) and streptomycin (0,1 mg/mL).
(iv) 1 mL of larvae suspension was added to 1 mL of the testing compound at a final concentration of 1,000 larvae/mL.
(v) Plates were incubated at 37°C in a 5% CO2 atmosphere.
(vi) Mortality rate was assessed through larvae motility after observation under optical microscopy for seven days. | [25] |
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| S. stercoralis | Compounds affecting L3 larvae mortality | (i) 50 L3 larvae in Locke’s nematode saline (LNS) suspension containing plant extract (50 mg/mL) and eosin (0.1 mg/mL) were kept in a light-free environment between counts.
(ii) The larvae were assessed as mobile, immobile (not stained), and dead (stained with eosin).
(iii) Relative activity (RA) was calculated by dividing LT50 of the most effective extract (shortest time to kill 50%) by the LT50 of each extract. | [26] |
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