Molluscicidal and antischistosomal activities of methanol extracts and isolated compounds from Eucalyptus globulus and Melaleuca styphelioides

Figures & data

Figure 1. Structures of the isolated compounds.

Table 1. Identification of the isolated compounds 1–14 using UV and HRESI/MS/MS.

No.	Isolated compounds	UV	(M-H)^−	Fragments (MS/MS)	References
1	Isobiflorin = 5,7-dihydroxy-2-methylchromone 8-C-β-d-glucopyranoside	208, 227sh, 256, 292, 320sh	353.08	233.04, 205.05	Tanaka et al. (1993)
2	5,7-Dihydroxy-2-isopropylchromone 8-C-β-d-glucopyranoside	208, 227sh, 253, 292, 320sh	381.12	261.07, 233.08, 218.05, 217.05	Wu et al. (1998)
3	Mallophenol B = 6S,9R-ionone 9-O-(6′-O-galloyl)-β-d-glucopyranoside	202, 218, 251, 291, 324	537.19	313.05, 271.04, 211.02, 169.01	Matsunami et al. (2006)
4	Eucalmainoside C = 2,4,6-trihydroxy-3-methyl-benzaldehyde  2-O-β-d-glucopyranoside	209, 229sh, 283, 320sh	329.08	167.03, 165.01, 139.04	Al-Sayed et al. (2010)
5	2,2,8-trimethyl-6-formyl-chrom-3-ene 7-O-β-d-glucopyranoside	202, 267, 315	395.13	233.08, 205.08	Ito et al. (2000)
6	Pedunculagin = 2,3-4,6-di-O-HHDP-(α/β)-d-glucopyranose	218, 273 (sh)	783.07	481.06, 301.00, 275.02	Hatano et al. (1988)
7	Tellimagrandin I = 2,3-di-O-galloyl-4,6-O-HHDP-β-d-glucopyranose	214, 271	785.08	301.00, 275.02, 169.01	Hatano et al. (1988)
8	Quercetin 3-O-α-l-arabinopyranoside-2′′-gallate	205, 264, 351	585.09	301.03, 273.04, 179.00, 151.01	Okamura et al. (1993)
9	Cornusiin B	214, 258, 357	1085.08	301.00	Okuda et al. (1984)
10	Benzyl alcohol 7-O-(3′,4′,6′-tri-O-galloyl)-β-d-glucopyranoside	222, 273	725.14	573.12, 421.11, 313.05, 169.01	Al-Sayed et al. (2010)
11	1,2,3,4,6-Penta-O-galloyl-β-d-glucopyranose	216, 275	939.11 (M-2H)^2−:469.05	617.08, 465.06, 169.01	Duan et al. (2004)
12	Eucalbanin B	214, 262	1569.16 (M-2H)^2−:784.08	765.05, 633.07, 301.00, 169.02	Yoshida et al. (1992)
13	Kaempferol 3-O-α-l-rhamnopyranoside	210, 262, 341	431.10	285.04	Mabry et al. (1970)
14	Casuarinin	216, 271	935.08 (M-2H)^2−:467.03	783.07, 633.07, 301.00, 169.01	Yoshida et al. (2008)

¹H and ¹³C NMR spectral data were in accordance with the previously reported data in the literature.

Table 2. Molluscicidial activity of extracts and isolated compounds on B. alexandrina snails.

Tested sample	LC10 (ppm)	LC25 (ppm)	LC50 (ppm)	LC90 (ppm)	Slope value
E. globulus extract	85.8	123.6	165.6	245.5	1.5
M. styphelioides extract	145.2	296.4	464.3	783.4	1.9
3	No effect
4	Not tested
5	174.5	343.4	531.1	887.8	1.9
6	85.5	185.6	297.0	508.4	2.0
7	17.4	65.7	119.5	221.5	2.5
8	No effect
9	59.3	108.6	163.4	267.5	1.8
10	Not tested
11	15.6	61.3	112.1	208.6	2.5
12	20.5	30.8	55.0	89.6	1.8
13	88.2	252.1	434.2	780.2	2.2
14	68.6	113.5	232.0	457.2	3.2
Niclosamide	0.1	0.2	0.3	0.4	1.9

Data represent the mortality rate of three separate experiments, each of the 10 snails.

Table 3. Changes in the biochemical parameters in hemolymph and soft tissues of B. alexandrina snails after 3 weeks exposure by the LC₂₅ of eucalbanin B (12).

	Hemolymph			Tissue
	Control	Exposed	% Change	Control	Exposed	% Change
ALP	74.1 ± 2.0	102.6 ± 0.8***	38.5	26.3 ± 2.3	74.0 ± 0.1***	181.4
ACP	546.0 ± 18.0	810.2 ± 3.6***	48.4	681.3 ± 107.9	825.9 ± 156.7**	21.2
AST	61.3 ± 0.7	130.7 ± 0.7***	113.2	350.0 ± 36.9	180.0 ± 10.0***	−48.6
ALT	50.7 ± 0.7	80.0 ± 0.6***	57.8	180.0 ± 10.0	166.7 ± 3.3**	−7.4
Total protein	3.1 ± 0.2	2.3 ± 0.1***	−25.8	2.2 ± 0.0	1.7 ± 0.3**	−22.7

Data represent the mean ± S.E. of three separate experiments. Test concentration = 30.8 ppm. Enzymes were expressed as units/g. Total protein as µg/mg.

**Significant at p < 0.01. ***Significant at p < 0.001.

Table 4. Effect of extracts and isolated compounds on Schistosoma mansoni miracidia, cercariae and worms after 1 and 2 h exposure.

	Miracidicidal activity (%)		Cercaricidal activity (%)		Schistosomicidal activity (%)
Tested sample	1 h	2 h	1 h	2 h	1 h	2 h
Control	0	5.0 ± 0	0	2.0 ± 0	0	2.0 ± 0
E. globulus extract	60.7 ±  0.3***	79.7 ± 0.3***	33.3 ± 0.3***	100 ± 0***	0	40.0 ± 0.6***
M. styphelioides extract	10.7 ± 0.3***	23.0 ± 0.6***	1.0 ± 0	22.3 ± 1.4***	0	0
3	99.7 ± 0.3***	99.7 ± 0.3***	5.7 ± 0.6**	41.0 ± 0.6***	7.0 ± 2.1 **	14.3 ± 3.5***
4	17.0 ± 0.6***	30.0 ± 0.6***	5.7 ± 0.7**	32.0 ± 1.5***	2.0 ± 0.6	12.3 ± 1.3**
5	62.0 ± 0.6***	99.7 ± 0.3***	13.0 ± 0.6***	40.0 ± 0.6***	9.3 ± 0.7***	13.0 ± 1.5**
6	62.0 ± 0.6***	80.0 ± 0.6***	0	14.0 ± 0.6***	1.0 ± 0	2.0 ± 0.6
7	67.0 ± 0.6***	80.0 ± 0.6***	8.4 ± 0.9***	38.3 ± 0.9***	7.3 ± 1.2**	19.3 ± 0.7***
8	0	7.0 ± 0.6	21.7 ± 4.4***	47.7 ± 3.9***	5.3 ± 0.9**	17.7 ± 1.4***
9	29.7 ± 0.3***	68.3 ± 0.3***	2.0 ± 0.6	40.0 ± 0.6***	0	13.0 ± 0.6**
10	66.7 ± 0.3***	99.7 ± 0.3***	11.0 ± 0.6***	45.0 ± 0.6***	12.0 ± 1.2***	25.7 ± 2.3***
11	7.0 ± 0.6**	13.0 ± 0.6	1.0 ± 0	22.0 ± 1.2***	0	5.7 ± 1.2
12	0	36.0 ± 0.6***	9.0 ± 0.4***	50.3 ± 0.6***	8.3 ± 0.9***	17.0 ± 0.6***
13	0	4.7 ± 0.9	16.7 ± 4.4***	50.0 ± 5.8***	0	7.3 ± 1.4
14	19.0 ± 0.6***	34.0 ± 0.6***	1.0 ± 0	8.0 ± 0	0	0
Praziquantal					ND	43.0 ± 1.5***

Data represent the % mortality ± SE of three separate experiments, each of the 100 organisms. Concentration of each tested sample is 20 ppm.

***Significant at p < 0.001. **Significant at p < 0.01.

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