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

Hazarika, S.; Barua, C. C.; Mohan, P.; Das, A.; Hazorika, M.; Tamuly, S.; Pathak, P.; Borah, B.; Phangchupi, D.; Chakraborty, A.; George, S.

doi:10.2478/helm-2025-0008

Figures & Tables

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.

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

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

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

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

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

Qualitative phytochemical analysis of HESEZA_

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

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

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

Fig. 1.

Fig. 2.

Fig. 3.

Fig. 4.

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

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

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

In vitro anthelmintic activity of different concentration of HESEZA against P_ cervi_

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

Qualitative phytochemical analysis of HESEZA_

In vitro effect of different concentration of HESEZA on various biochemical parameters following 4 hours of incubation of adult P_ cervi_

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