In vitro and in vivo anthelmintic response of the seeds of Amomum subulatum roxb and Vitex negundo

Badar, S. N.; Sajid, M. S.; Rizwan, H. M.; Abbas, H.; Maqbool, M.; Malik, M. A.; Nisar, R. H. A.; Rajput, M.; Shamim, A.; Akram, Q.; Naeem, M. A.; Alvi, M. A.; Iqbal, Z.; Hassan, M. A.; Ghazanfer, S.; Iqbal, Z.

doi:10.1590/1519-6984.261768

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Brazilian Journal of Biology

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Original Article • Braz. J. Biol. 84 • 2024 • https://doi.org/10.1590/1519-6984.261768 copy

In vitro and in vivo anthelmintic response of the seeds of Amomum subulatum roxb and Vitex negundo

Atividade anti-helmíntica in vitro e in vivo das sementes de Amomum subulatum roxb e Vitex negundo

Authorship SCIMAGO INSTITUTIONS RANKINGS

Abstract

The current study was designed to check the anthelmintic activities of some local plants. Seeds of Amomum (A.) subulatum and Vitex (V.) negundo in different solvents were subjected to in vitro (adult motility assay; AMA and egg hatch assay; EHA) and in vivo (faecal egg count reduction test; FECRT) anthelmintic activity testing protocols using Haemonchus (H.) contortus as an experimental model. The results of AMA, EHA, and FECRT were statistically analysed through linear regression and Duncan multiple range test. In AMA test, at 50 mg mL^-1 concentration, the percent mortality of H. contortus was higher in A. subulatum than V. negundo, whereas, in EHA test, A. subulatum was proven better ovicidal (LC₅₀=14.2 µg mL^-1) than V. negundo (LC₅₀= 65.7405 µg mL^-1). The FECRT also indicated the better efficacy of A. subulatum than V. negundo against natural infection of gastrointestinal (GI) parasites. The crude powder of plants used in this study showed 29.6% to 57.7% anthelmintic. The reduction rate was found higher for A. subulatum (3 g kg^-1) as compared to V. negundo (7 g kg^-1). Reagrding efficacy analysis of solvents used for plants extract, ethyl acetate and chloroform were found better in increasing ovicidal activity in adult worms (in vitro testing), whereas, the crude aqueous methanol was found better than the crude powders in in vivo testing. It will be beneficial to document the indigenous knowledge to standard scientific procedures for their validation. This study will help to motivate the farmers to make a better choice of cultivation of the indigenous plants because of their varying efficacies as an alternative preventive approach against the GI parasitic infections.

Keywords:
Amomum subulatum; Vitex negundo; Haemonchus contortus; In vivo and in vitro assay

Group	Treatment	Dose rate
1	untreated or control
2	levamisole	7.5 mg per kg b.wt.
3	crude powder (CP) of A. subulatum (S.)	0.5 g per kg b.wt.
4	crude powder (CP) of A. subulatum (S.)	1 g per kg b.wt.
5	crude powder (CP) of A. subulatum (S.)	3 g per kg b.wt.
6	crude powder (CP) of V. negundo (S.)	1 g per kg b.wt.
7	crude powder (CP) of V. negundo (S.)	4 g per kg b.wt.
8	crude powder (CP) of V. negundo (S.)	7 g per kg b.wt.
9	^* * CAME = Crude aqueous methanol extract. CAME of A. subulatum (S.)	0.5 g per kg b.wt.
10	CAME of A. subulatum (S.)	1 g per kg b.wt.
11	CAME of A. subulatum (S.)	3 g per kg b.wt.
12	CAME of V. negundo (S.)	1 g per kg b.wt.
13	CAME of V. negundo (S.)	4 g per kg b.wt.
14	CAME of V. negundo (S.)	7 g per kg b.wt.

Treatments mg mL^-1	Amomum subulatum (Seeds)				Vitex negundo (Seeds)
Treatments mg mL^-1	Mean±SE of dead worms at different hours				Mean±SE of dead worms at different hours
Hours post-exposure	0 hour	2 hours	6 hours	10 hours	0 hour	2 hours	6 hours	10 hours
Levamisole 0.5	0^r	10.00±0.00^a	10.00±0.00^a	10.00±0.00^a	0^k	10.00±0.00^a	10.00±0.00^a	10.00±0.00^a
PBS	0^r	0^r	0^r	0^r	0^k	0^k	0^k	0^k
Crude Aqueous Methanolic Extract
1.56	0^m	0^m	0.33±0.33^lm	1.00±0.00 ^klm	0^o	0^o	0^k	0.67±0.67^mno
3.12	0^m	0^m	1.00±0.00^klm	1.67±0.00^ijk	0^o	0^o	0.33±0.33^ijk	1.00±0.58^lo
6.25	0^m	0^m	1.33±0.33^jkl	2.33±0.33^hij	0^o	0^o	0.67±0.33^hk	3.00±0.58^fi
12.5	0^m	0^m	1.67±0.33^ijk	3.33±0.33^h	0^o	0^o	1.67±0.33^gj	4.00±0.58^ef
25	0^m	1.67±0.33^ijk	3.00±0.58^h	6.00±0.58^def	0^o	1.00±0.58^lo	2.33±0.33^eh	4.33±0.33^de
50	0^m	2.67±0.88^hi	5.67±0.33^efg	7.33±0.67^bc	0^o	2.00±0.58^il	4.00±0.58^cde	7.00±0.58^c
Chloroform fractions
1.56	0^r	0^r	0.67±0.67^qr	1.67±0.33^nq	0^k	0^k	0^k	0.33±0.33^jk
3.12	0^r	0^r	1.33±0.33^or	2.67±0.33^ko	0^k	0^k	0.67±0.33^ijk	1.33±0.33^gj
6.25	0^r	0.67±0.33^qr	2.00±0.00^mq	3.00±0.00^jn	0^k	0^k	1.00±0.00^hk	1.67±0.33^fi
12.5	0^r	1.33±0.33^or	2.67±0.67^ko	4.00±0.58^gk	0^k	0^k	1.33±0.33^gj	2.33±0.33^dg
25	0^r	1.67±0.33^nq	4.33±0.67^fj	5.33±0.67^efg	0^k	0.33±033^jk	2.00±0.58^eh	3.00±0.58^cde
50	0^r	3.33±0.88^im	5.67±0.88^ef	7.67±0.33^bc	0^k	1.00±0.58^hk	3.00±0.58^cde	4.67±0.67^b
Ethyl acetate fractions
1.56	0^r	0^r	1.33±0.33^opq	2.67±0.33^kn	0^m	0^m	0^m	0.33±0.33^lm
3.12	0^r	0.67±0.33^qr	2.33±0.33^lo	3.33±0.67^jkl	0^m	0^m	0.67±0.33^klm	1.33±0.33^ijk
6.25	0^r	1.00±0.00^pqr	2.67±0.33^kn	4.33±0.88^hij	0 ^m	0^m	1.33±0.33^ijk	2.00±0.00^ghi
12.5	0^r	1.67±0.33^nq	4.33±0.33^hij	6.00±0.58^ef	0^m	0^m	1.67±0.33^hij	2.33±0.33^gh
25	0^r	2.00±0.58^mp	5.33±0.33^fgh	9.00±0.58^ab	0^m	0.67±0.33^klm	2.00±0.00 ^ghi	4.00±0.58^de
50	0^r	4.33±0.33^hij	7.00±0.58^de	10.00±0.00^a	0^m	1.67±0.33^hij	3.33±0.88 ^ef	6.00±0.58^b
Petroleum spirit fractions
1.56	0ⁿ	0ⁿ	0ⁿ	1.00±0.00^klm	0^k	0^k	0^k	0.33±0.33^jk
3.12	0ⁿ	0ⁿ	0ⁿ	1.67±0.33^ijk	0^k	0^k	0^k	0.67±0.33^ijk
6.25	0ⁿ	0ⁿ	0.67±0.33^lmn	2.33±0.33^ghi	0^k	0^k	0^k	1.33±0.33^ghi
12.5	0ⁿ	0ⁿ	1.33±0.33^jkl	2.67±0.33^fgh	0^k	0^k	0.67±0.33^ijk	2.00±0.58^efg
25	0ⁿ	0ⁿ	2.00±0.58^hij	3.33±0.33^def	0^k	0^k	1.33±0.33^ghi	2.67±0.33^de
50	0ⁿ	0ⁿ	3.00±0.58^efg	4.67±0.67^bc	0^k	0^k	2.67±0.67^de	4.33±0.33^b
Aqueous fractions
1.56	0^k	0^k	0^k	0.33±0.33^jk	0^l	0^l	0^l	0.33±0.33^kl
3.12	0^k	0^k	0^k	0.67±0.33^ijk	0^l	0^l	0^l	1.33±0.33^ijk
6.25	0^k	0^k	0^k	1.33±0.33^ghi	0^l	0^l	0.67±0.33^jkl	1.67±0.33^hij
12.5	0^k	0^k	0^k	2.33±0.33^ef	0^l	0^l	1.00±0.58^ijkl	2.67±0.67^fgh
25	0^k	0^k	1.33±0.33^ghi	2.67±0.33^de	0^l	0.33±0.33^kl	1.67±0.33^hij	3.67±0.88^ef
50	0^k	0^k	2.33±0.33^ef	5.00±0.58^b	0^l	0.67±0.67^jkl	3.00±0.58^fg	5.67±0.67^bc

Treatments	fraction concentrations mg mL^-1					LC₅₀ µg mL^-1
	1.2	12	120	1200	12000
Albendazole	79	64	40	5	0	0.0345
Amomum subulatum (Seeds)
CAME	41	39	28	19	6	14.1773
Choloroform fraction	38	37	33	27	13	19.1514
Ethyl acetate fraction	45	37	28	17	4	19.6203
Aqueous fraction	44	42	33	23	17	21.4279
Petroleum spirit fraction	45	37	33	27	13	254.9207
Vitex negundo (Seeds)
CAME	77	66	46	31	12	65.7405
Aqueous fraction	73	61	40	34	14	39.7628
Petroleum spirit fraction	75	58	48	36	18	62.5137
Ethyl acetate fraction	77	66	50	39	17	93.3188
Choloroform fraction	89	67	49	36	26	192.0218

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