In vitro evaluation of hydro-ethanolic seed extract of Zanthoxyllum armatum DC to establish its mechanism of action for paralysis of Paramphistomum cervi in continuation with in vivo experimentation in naturally infected goat

SI. No	Active principle	Test applied	Result
1.	Alkaloid	Hager's test	Positive
2.	Terpenoids	Salkowski test	Positive
3.	Diterpenes	Copper acetate test	Positive
4.	Flavonoids	Ferric chloride test	Positive
5.	Steroid	Salkowski test	Positive
6.	Tannins	Ferric chloride test	Positive
7.	Glycosides	Sodium hydroxide test	Positive
8.	Saponins	Frothing test	Negative
9.	Phenolic compound	Ferric chloride test	Positive

Table 2.

In vitro anthelmintic activity of different concentration of HESEZA against P. cervi.

Drug/Extract	Conc (μg/ml)	No of parasite showing motility (in minutes)
		0	30	60	90	120	150	180	210	240	270	300
Control	-	6	6	6	6	6	6	6	6	6	6	6
Z. armatum (Hydro-ethanolic seed extract)	50	6	6	6	6	6	6	6	6	5	4	3
	150	6	6	6	6	6	5	5	4	3	2	1
	300	6	6	6	5	4	3	1	0	0	0	0
500	6	5	4	3	1	0	0	0	0	0	0
1000	6	4	3	1	1	0	0	0	0	0	0
3000	6	3	2	1	0	0	0	0	0	0	0
Oxyclozanide	10⁻⁵ M	6	2	0	0	0	0	0	0	0	0	0

Table 3.

In vitro effect of different concentration of HESEZA on various biochemical parameters following 4 hours of incubation of adult P. cervi.

Biochemical parameters	Control	Standard (Oxyclozanide 10⁻³ M)	Concentrations (μg/ml) of seed extract
Biochemical parameters	Control	Standard (Oxyclozanide 10⁻³ M)	100	300	1000	3000
Glucose uptake (mg/100mg of tissue)	1.52±0.05^a	1.08±0.03^e	1.45±0.07^a	1.34±0.07^b	1.19±0.09^c	1.09±0.08^d
Glycogen content (mg/100mg of tissue)	2.54±0.03^a	1.84±0.09^e	2.47±0.04^a	2.30±0.08^b	2.10±0.03^c	1.87±0.07^d
Lactic acid production (μg/g) of tissue	80.47±5.18^a	116.28±4.34^e	85.27±4.87^a	97.45±6.40^b	108.16±4.18^c	116.45±3.27^e
Acetylcholinesterase activity (mM of Ach hydrolyzed/g weight/h)	92.09±5.70^a	59.17±4.48^e	91.00±4.18^a	88.55±4.89^ab	83.00±4.19^ab	70.64±3.66^c

[i] Values are the Mean ± S.E. for six replicates.

[ii] Mean value in each row bearing common superscripts do not differ significantly at P<0.05.

Effect of different concentration of HESEZA on glucose uptake in adult *P. cervi*.

Effect of different concentration of HESEZA on glycogen content in adult *P. cervi*.

Effect of different concentration of HESEZA on lactic acid production in adult *P. cervi*.

Effect of different concentration of HESEZA on acetylcholinesterase activity in adult *P. cervi*.

Table 4.

In vivo study of HESEZA (150 and 250 mg/kg b. wt) on EPG in experimental groups at different days of treatment.

Days of observation
Group	0^th	7^th	14^th	21^st	28^th
A	585±59.15^a	591.67±63.21^a	601.67±64^a	586.67±59.48^a	573.33±45.07^a
B	639±33.64^a	470±18.26^ab	326.33±12.33^bc	151.5±13.58^c	20.33±3.87^d
C	603.33±32.52^a	389.67±47.14^b	154.83±40.63^c	46.5±19.19^d	0±0
D	516.67±31.16^a	121.33±9.61^b	8±1.46^c	0±0	0±0

[i] Values are the Mean ± S.E. for six replicates.

[ii] Mean value in each row bearing common superscripts do not differ significantly at P<0.05 and P<0.01.

Table 5.

In vivo study of HESEZA (150 and 250 mg/kg b. wt) on body weight in experimental groups at different days of treatment.

Days of observation
Group	0^th	7^th	14^th	21^st	28^th
A	7.77±0.17^a	7.78±0.17^a	7.81±0.17^a	7.82±0.18^a	7.84±0.18^a
B	7.56±0.25^a	8.04±0.21^ab	8.61±0.23^ab	9.25±0.23^b	9.84±0.24^b
C	7.93±0.15^a	8.45±0.17^ab	9.11±0.2^b	9.85±0.21^b	10.88±0.26^bc
D	7.79±0.2^a	8.36±0.2^ab	9.32±0.3^b	10.26±0.19^bc	11.37±0.18^c

[i] Values are the Mean ± S.E. for six replicates.

[ii] Mean value in each row bearing common superscripts do not differ significantly at P<0.05.

Table 6.

In vivo study of HESEZA (150 and 250 mg/kg b. wt) on hematological parameters in experimental groups at different days of treatment.

Group	Parameter	Days of observation
		0^th	7^th	14^th	21^st	28^th
A	Hb (g/dl)	7±0.06^a	6.97±0.06^a	6.97±0.04^a	6.9±0.07^a	6.85±0.08^a
B		7.03±0.11^a	7.07±0.13^a	7.77±0.12^ab	8.02±0.12^c	8.28±0.11^c
C		7.1±0.1^a	7.17±0.08^a	7.32±0.08^a	7.83±0.1^ab	8.57±0.08^c
D		6.92±0.06^a	7.85±0.04^ab	8.05±0.04^c	8.6±0.04^c	9.25±0.08^d
A	PCV (%)	20.55±0.34^a	20.57±0.36^a	20.6±0.34^a	20.63±0.34^a	20.68±0.3^a
B		21.85±0.28^a	21.88±0.29^a	21.97±0.3^ab	22.03±0.28^b	22.13±0.29^b
C		20.48±0.34^a	21.53±0.32^a	21.87±0.32^a	22.08±0.32^b	22.57±0.32^b
D		20.32±0.31^a	21.78±0.27^a	22.05±0.26^b	22.77±0.29^b	22.93±0.29^b
A	TLC (10³/mm³)	15.78±0.4^a	15.86±0.43^a	16.24±0.21^a	16.17±0.2^a	16.09±0.29^a
B		15.47±0.25^a	14.72±0.35^ab	13.68±0.31^b	12.33±0.31^bc	11.35±0.33^c
C		15.68±0.22^a	14.1±0.3^ab	12.63±0.35^bc	11.27±0.37^c	9.72±0.32^d
D		15.29±0.3^a	12.15±0.36^bc	10.8±0.27^cd	9.38±0.26^d	8.9±0.24^e
A	TEC (10⁶/mm³)	10.48±0.17^a	10.28±0.27^a	10.34±0.2^a	10.41±0.22^a	10.38±0.19^a
B		9.62±0.18^a	10.33±0.15^a	11.05±0.19^ab	11.8±0.19^ab	12.55±0.16^b
C		10.2±0.28^a	11.23±0.3^ab	12.07±0.29^b	13.02±0.28^bc	13.78±0.26^c
D		10.31±0.32^a	12.08±0.12^b	13.29±0.21^bc	13.7±0.16^c	13.77±0.19^c
A	Neutrophil count (%)	51.4±0.36^a	51.35±0.33^a	51.68±0.38	51.79±0.45	51.58±0.45
B		52.63±1.03^a	48.74±0.67^b	44.33±0.83^bc	40.57±0.77^c	36.34±0.92^d
C		50.72±1.03^a	46.11±0.87^b	41.84±0.93^c	36.97±1.1^d	31.95±1.2^e
D		51.01±0.47^a	41.28±0.86^b	35.59±0.6^c	30.76±0.31^d	29.25±0.34^e
A	Eosinophil count (%)	5.62±0.21^a	5.63±0.2^a	5.66±0.2^a	5.69±0.2^a	5.7±0.22^a
B		5.11±0.13^a	4.53±0.14^b	3.88±0.1^c	3.19±0.11^c	2.42±0.12^d
C		5.6±0.28^a	4.92±0.32^b	4.18±0.33^b	3.52±0.31^c	2.65±0.26^d
D		5.51±0.17^a	4.6±0.17^b	3.83±0.17^c	3.05±0.17^c	2.25±0.15^d
A	Monocyte count (%)	4.08±0.13^a	4.08±0.12^a	4.08±0.13^a	4.13±0.14^a	4.1±0.13^a
B		4.17±0.09^a	3.92±0.09^a	3.55±0.14^b	3.24±0.14^b	2.9±0.11^bc
C		3.98±0.12^a	3.64±0.13^ab	3.24±0.11^b	2.88±0.1^bc	2.36±0.11^c
D		4.1±0.12^a	3.47±0.14^b	3.05±0.11^bc	2.62±0.16^c	2.29±0.14^c
A	Lymphocyte count (%)	35.17±1.35^a	35.04±1.14^a	35.41±1.29^a	35.05±1.35^a	35.21±1.28^a
B		34.17±1.07^a	38.21±1.06^b	42.26±1.04^c	46.8±0.82^cd	51.37±0.63^e
C		35.62±0.93^a	40.11±0.93^bc	45.04±0.89^c	49.77±0.88^d	54.91±0.8^e
D		35.74±0.7^a	42.72±1.11^bc	48.92±1.16^c	55.24±1.39^d	60.8±1.27^e

[i] Values are the Mean ± S.E. for six replicates.

[ii] Mean bearing different superscripts in column differ significantly (P<0.05; P<0.01) in groups.

Table 7.

In vivo study of HESEZA (150 and 250 mg/kg b. wt) on biochemical parameters in experimental groups at different days of treatment.

Group	Parameter	Days of observation
		0^th	7^th	14^th	21^st	28^th
A	Total serum protein (g/dl)	4.28±0.11^a	4.24±0.09^a	4.26±0.1^a	4.25±0.1^a	4.23±0.09^a
B		4.27±0.09^a	4.67±0.11^a	5.03±0.17^ab	5.37±0.17^ab	5.79±0.16^b
C		4.31±0.09^a	4.77±0.07^a	5.21±0.13^ab	5.62±0.13^b	6.03±0.14^bc
D		4.38±0.08^a	4.84±0.13^a	5.23±0.14^ab	5.65±0.14^b	6.19±0.1^bc
A	Serum albumin (g/dl)	1.82±0.04^a	1.81±0.04^a	1.81±0.03^a	1.81±0.03^a	1.82±0.04^a
B		1.75±0.03^a	1.96±0.05^a	2.26±0.08^ab	2.64±0.08^b	2.94±0.06^bc
C		1.83±0.03^a	2.21±0.08^ab	2.64±0.11^b	2.97±0.09^bc	3.24±0.09^c
D		1.8±0.04^a	2.19±0.09^ab	2.64±0.08^b	2.99±0.08^bc	3.26±0.08^c
A	Serum globulin (g/dl)	2.27±0.05^a	2.24±0.05^a	2.25±0.05^a	2.25±0.06^a	2.26±0.06^a
B		2.16±0.08^a	2.35±0.06^ab	2.56±0.07^b	2.77±0.06^c	2.94±0.05^d
C		2.24±0.09^a	2.45±0.08^ab	2.66±0.07^b	2.84±0.06^c	3.03±0.06^d
D		2.15±0.07^a	2.39±0.07^ab	2.59±0.08^b	2.85±0.06^c	3.06±0.06^d
A	Aspartate aminotransferase level (U/L)	102.65±2.04^a	102.95±1.82^a	103.32±1.7^a	103.32±1.69^a	103.7±1.51^a
B		100.89±1.51^a	93.28±1.69^ab	85.82±1.56^b	78.27±1.64^bc	70.29±1.43^c
C		102.99±1.46^a	92.93±1.24^ab	84.92±1.37^b	76.84±1.04^bc	67.62±1.14^c
D		103.75±2.16^a	90.85±1.82^ab	82.07±1.95^b	71.54±1.22^bc	62.49±1.11^cd
A	Alanine aminotransferase level (U/L)	27.81±0.6^a	27.8±0.61^a	28.21±0.57^a	28.22±0.53^a	28.39±0.57^a
B		26±0.7^a	24.85±0.62^b	23.86±0.55^c	21.9±0.39^d	20.54±0.44^d
C		27.23±0.38^a	25.5±0.54^b	23.71±0.4^c	22.12±0.36^cd	19.83±0.6^de
D		27.38±0.55^a	25.47±0.62^b	23.02±0.43^c	20.57±0.55^d	18.96±0.36^e
A	Alkaline phosphatase level (U/L)	189.72±1.17^a	190.44±1.36^a	191.76±1.5^a	192.05±1.19^a	192.5±1.2^a
B		189.38±1.42^a	177.65±1.31^ab	167.06±1.69^bc	156.64±1.74^cd	145.1±1.47^d
C		188.25±1.43^a	176.9±1.77^ab	164.18±2.28^c	152.66±1.84^d	140.79±2.2^e
D		189.1±2.08^a	174.96±1.76^b	161.76±1.48^c	148.04±1.03^d	136.47±1.22^e
A	Serum iron (µg/dl)	150.26±1.42^a	150.32±1.42^a	150.29±1.43^a	150.26±1.44^a	150.25±1.43^a
B		152.07±1.54^a	155.1±1.5^ab	159.68±1.63^b	163.59±1.6^c	167.52±1.37^cd
C		152.52±1.3^a	158.22±1.67^b	163.05±1.63^c	168.7±1.75^cd	173.5±1.73^d
D		152.2±1.1^a	157.53±1.15^b	162.54±1.17^c	167.55±0.86^cd	172.64±0.81^d
A	Serum copper (µg/dl)	61.99±0.88^a	61.83±0.82^a	61.93±0.83^a	61.99±0.84^a	62.03±0.84^a
B		62.39±1.48^a	65.17±1.51^ab	68.11±1.46^b	71.48±1.16^bc	75.38±1.28^c
C		62.63±1.5^a	66.94±1.2^ab	71.95±1.29^bc	76.16±1.31^c	80.61±1.05^cd
D		60.88±1.38^a	66.86±1.13^ab	72.33±1.15^bc	77.77±0.85^c	83.54±0.8^cd
A	Serum sodium level (mmol/L)	124.06±1.4^a	123.67±1.31^a	123.5±1.21^a	123.72±1.16^a	123.84±1.13^a
B		125.93±0.9^a	127.79±0.94^ab	129.5±0.95^b	131.5±0.97^bc	133.18±0.8^bc
C		123.51±1.18^a	125.91±0.98^a	128.53±1.01^ab	131.09±0.94^bc	134.1±1.05^c
D		124.01±1.31^a	128.83±1.38^ab	132.05±1.53^bc	135.62±1.58^c	139.14±1.52^d
A	Serum potassium level (mmol/L)	3.86±0.15^a	3.84±0.15^a	3.85±0.15^a	3.86±0.15^a	3.87±0.15^a
B		3.22±0.17^a	4.37±0.15^bc	4.5±0.15^c	4.58±0.15^cd	4.69±0.15^d
C		3.59±0.15^a	4.01±0.08^ab	4.24±0.07^b	4.52±0.09^cd	4.75±0.07^d
D		3.92±0.09^a	4.29±0.13^b	4.5±0.11^cd	4.69±0.09^d	4.93±0.09^e
A	Serum chloride level (mmol/L)	69.79±0.95^a	69.68±0.97^a	69.77±0.99^a	69.87±0.98^a	70.02±0.93^a
B		71.27±1.89^a	74.41±1.92^b	76.75±2.01^bc	80.27±2.24^c	84.92±2.34^d
C		71.09±2.09^a	74.76±1.94^b	79.02±2.07^c	84.16±1.51^d	88.91±1.12^e
D		69.37±1.64^a	74.95±1.69^b	81±1.78^cd	88.67±2.24^e	96.55±2.08^f

[i] Values are the Mean ± S.E. for six replicates.

[ii] Mean bearing different superscripts in column differ significantly (P<0.05; P<0.01) in groups.

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DOI: https://doi.org/10.2478/helm-2025-0008 | Journal eISSN: 1336-9083 | Journal ISSN: 0440-6605

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Language: English

Page range: 19 - 29

Submitted on: Jul 16, 2024

Accepted on: Apr 15, 2025

Published on: May 24, 2025

Published by: Slovak Academy of Sciences, Institute of Parasitology

In partnership with: Paradigm Publishing Services

Publication frequency: Volume open

Keywords:

Zanthoxyllum armatum DC,

Paramphistomum cervi,

Related subjects:

Zoology,

Ecology,

Life sciences, other,

Medicine,

Clinical medicine,

Microbiology, virology and infection epidemiology

© 2025 S. Hazarika, C. C. Barua, P. Mohan, A. Das, M. Hazorika, S. Tamuly, P. Pathak, B. Borah, D. Phangchupi, A. Chakraborty, S. George, published by Slovak Academy of Sciences, Institute of Parasitology
This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 License.

Volume 62 (2025): Issue 1 (March 2025)

In vitro evaluation of hydro-ethanolic seed extract of Zanthoxyllum armatum DC to establish its mechanism of action for paralysis of Paramphistomum cervi in continuation with in vivo experimentation in naturally infected goat

Figures & Tables

Table 1.

Table 2.

Table 3.

Fig. 1.

Fig. 2.

Fig. 3.

Fig. 4.

Table 4.

Table 5.

Table 6.

Table 7.

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